Heat stress impairs oocyte maturation through ceramide-mediated apoptosis in pigs
Heat stress (HS) is an emerging issue that greatly impairs the reproductive performance of animals and humans. In particular, disruption of oocyte maturation due to HS is considered a major cause of impaired reproductive performance. HS is known to induce ceramide generation, which causes reactive oxygen species (ROS) production and mitochondrial dysfunction, thereby inducing apoptosis. Therefore, we investigated whether inhibition of ceramide generation ameliorates HS-induced apoptosis in porcine cumulus-oocyte complexes (COCs) using specific inhibitors of the de novo (fumonisin B1, FB1) and hydrolytic pathways (desipramine, Des) of ceramide formation. We investigated the effects of FB1 and Des supplementation under HS conditions (41.5 °C for 44 h) on in vitro maturation (IVM) of porcine COCs. After IVM, HS significantly reduced proportion of COCs exhibiting fully expanded cumulus cells and the rate of metaphase II in oocytes. After parthenogenetic activation (PA), HS significantly reduced the rates of cleavage and blastocyst formation with a lower total cell number and a higher percentage of apoptosis in blastocysts.
However, FB1 or Des supplementation under HS avoided detrimental effects of HS on expansion of cumulus cells, nuclear maturation of oocytes, and embryonic development after PA including the rates of cleavage and blastocyst formation, total cell number, and the percentage of apoptosis in blastocysts. Furthermore, FB1 or Des addition under HS, compared with HS alone, significantly decreased ceramide generation, ROS production, cytochrome C expression, and apoptosis and increased mitochondrial membrane potential in COCs, reaching levels comparable with those of the control. Taken together, our results indicate that HS impaired oocyte maturation through ceramide-mediated apoptosis.
Finally, the interaction of SCGN with ATF4 was computationally predicted and then validated by a co-immunoprecipitation assay. We found that ox-LDL treatment increased the levels of ER stress markers, such as phosphorylated protein kinase-like endoplasmic reticulum kinase, phosphorylated eukaryotic initiation factor 2 alpha, activating transcription factor 4 (ATF4), and transcription factor CCAAT-enhancer-binding protein homologous protein, and promoted MIN6 cell apoptosis; in addition, the expression of SCGN was downregulated. siRNA-mediated SCGN knockdown exacerbated β-cell ER stress by increasing ATF4 expression.
Tetrahedral Framework Nucleic Acid Inhibits Chondrocyte Apoptosis and Oxidative Stress through Activation of Autophagy
Osteoarthritis (OA) is a degenerative articular cartilage pathogenic process that is accompanied by excessive chondrocyte apoptosis. The occurrence of chondrocyte death and OA is related to decreased autophagy. Tetrahedral framework nucleic acid (TFNA), a potent bioactive DNA nanomaterial, exerts antiapoptotic and antioxidative effects in various diseases, resulting in autophagy promotion and inhibition of the Wnt/β-catenin-signaling pathway. Here, we aimed to elucidate the therapeutic effects of TFNA on OA and its potential molecular mechanism of action. TFNA was synthesized and characterized by established methods. An interleukin (IL)-1β stimulated OA cell model was established and treated with TFNA. Cellular uptake of TFNA and intracellular reactive oxygen species levels were examined via immunofluorescence and flow cytometry.
Apoptotic cell death was documented by the Cell Counting Kit-8 (CCK8) assay and flow cytometry. Transmission electron microscopy was applied to view the autophagosomes. The expression of BCL2, BAX, caspase-3, Nrf2, HO-1, LC3-II, Beclin1, Atg7, β-catenin, Lef-1, and CyclinD1 was detected by immunofluorescence and western blotting. TFNA was successfully synthesized and effectively entered chondrocytes in the absence or presence of IL-1β without the help of transfection agents. TFNA treatment in IL-1β-induced chondrocytes reduced apoptosis by activating the BCL2/BAX/caspase-3 pathway, inhibited oxidative stress by regulating the Nrf2/HO-1-signaling pathway, and enhanced autophagy through upregulated LC3-II, Beclin1, and Atg7. Moreover, TFNA showed chondroprotective effects by regulating the Wnt/β-catenin-signaling pathway. Overall, TFNA may have utility as a therapeutic nanomedicine for OA.
Heat stress impairs oocyte maturation through ceramide-mediated apoptosis in pigs
Raddeanin A suppresses lung cancer cell proliferation via induction of apoptosis and increased production of ROS
At present, in vitro cell experiments have confirmed that RaddeaninA can effectively inhibit the proliferation of some tumor cells, but the effect of RaddeaninA on lung cancer cells has not been observed. Therefore, this study explored its effect on lung cancer cells and its mechanism of action. Human lung cancer cell lines were treated with serum-free medium and varied concentrations of Raddeanin A. Cell proliferation and apoptosis were determined using MTT, and flow cytometric assays, respectively. The intracellular level of ROS was determined using DCFH-DA assay. Protein and mRNA expressions of bax, bcl-2 and cyt c were measured using Western blotting and qRT-PCR. RaddeaninA treatment can promote PC-9 cell apoptosis in a time and dose-dependent manner (p<0.05).
Treatment of PC-9 cells with Raddeanin significantly and dose-dependently increased the activities of caspase-9 and caspase-3 (p<0.05), and led to significant and dose-dependent increases in ROS levels. Treatment of PC-9 cells with Raddeanin A led to significant and dose-dependent decreases in mitochondrial membrane potential. It significantly and dose-dependently upregulated bax mRNA and protein expressions, but down-regulated bcl-2 mRNA and protein expressions significantly and dose-dependently (p<0.05). On the other hand, Raddeanin significantly and dose-dependently down-regulated cytoplasmic bax protein expression, while upregulating cyt c expression (p<0.05).
Description: Anti-Kappa recognizes surface immunoglobulin on normal and neoplastic B-cells, and has been indicated as a potential aid in the identification of leukemias, plasmacytomas, and certain non-Hodgkin's lymphomas, where the expression of a single light chain class is restricted. The determination of light chain ratio is critical in evaluating B-cell neoplasms, as the majority of B-cell lymphomas express either kappa or lambda light chains, while a mixture of kappa and lambda is characteristic of reactive proliferations. In paraffin-embedded tissue, Anti-Kappa displays strong staining of kappa-positive plasma cells, as well as cells that have absorbed exogenous immunoglobulins.
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-GOT2 (aa 295 to 306) . This antibody is tested and proven to work in the following applications:
Polyclonal Goat Anti-GOT2 (aa 360 to 373) Antibody
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-GOT2 (aa 360 to 373) . This antibody is tested and proven to work in the following applications:
Polyclonal Goat Anti-Renalase (aa 134 to 147) Antibody
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-Renalase (aa 134 to 147) . This antibody is tested and proven to work in the following applications:
Polyclonal Goat Anti-Renalase (aa 224 to 233) Antibody
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-Renalase (aa 224 to 233) . This antibody is tested and proven to work in the following applications:
Description: IkappaB-alpha is a protein encoded by the NFKBIA gene which is approximately 35,6 kDa. IkappaB-alpha is localised to the cytoplasm and nucleus. It is involved in activated TLR4 signalling, the TNFR1 pathway, 4-1BB pathway and toll-like receptor signalling pathways. This protein falls under the NF-kappa-B inhibitor family. It inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses it becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription. IkappaB-alpha is expressed in adherent monocytes. Mutations in the NFKBIA gene may result in ectodermal dysplasia and leukorrhea. STJ93782 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of IkappaB-alpha protein.
Description: IkappaB beta is a protein encoded by the NFKBIB gene which is approximately 37,7 kDa. IkappaB beta is localised to the cytoplasm and nucleus. It is involved in activated TLR4 signalling, toll-like receptor signalling pathways and the 4-1BB pathway. This protein falls under the NF-kappa-B inhibitor family, which inhibits NF-kappa-B by complexing with it and trapping it in the cytoplasm. Phosphorylation of serine residues on the protein marks it for destruction via the ubiquitination pathway, thereby allowing activation of the NF-kappa-B, which then translocates to the nucleus to function as a transcription factor. IkappaB beta is expressed in the liver, lung, pancreas, nervous system and blood. STJ97539 was developed from clone 1F3 and was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This primary antibody detects endogenous levels of IkappaB beta.
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-BDH2 / DHRS6 (aa 60 to 71) . This antibody is tested and proven to work in the following applications:
Polyclonal Goat Anti-EPB41L2 / 4.1G (aa 593 to 604) Antibody
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-EPB41L2 / 4.1G (aa 593 to 604) . This antibody is tested and proven to work in the following applications:
Polyclonal Goat Anti-TUSC4 / NPRL2 (aa 140 to 151) Antibody
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-TUSC4 / NPRL2 (aa 140 to 151) . This antibody is tested and proven to work in the following applications:
Polyclonal Goat Anti-C2GnT-M (aa 273 to 284) Antibody
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-C2GnT-M (aa 273 to 284) . This antibody is tested and proven to work in the following applications:
Polyclonal Goat Anti-C2GnT-M (aa 410 to 422) Antibody
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-C2GnT-M (aa 410 to 422) . This antibody is tested and proven to work in the following applications:
Should the Human Ghrelin-O-Acyltransferase (GOAT) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
Description: A sandwich quantitative ELISA assay kit for detection of Human Ghrelin-O-Acyltransferase (GOAT) in samples from serum, plasma, tissue homogenates or other biological fluids.
Should the Human Ghrelin-O-Acyltransferase (GOAT) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
Description: A sandwich quantitative ELISA assay kit for detection of Human Ghrelin-O-Acyltransferase (GOAT) in samples from serum, plasma, tissue homogenates or other biological fluids.
What are the applications and the optimal dilutions of the product Antibody against PARP (Clone 10H)for analysis? - Immunocytochemistry : 5-20 ?g/ml | Western Blot (10 ?g/ml using colorimetric methods | <2 ?g/ml for ECL) | Immunoprecipitation and ELI
Description: Goat IgG Secondary antibody Clone: pAb against UltraPolymer Goat Anti-Human IgG (H&L) HRPfor application in IHC.The immunogen is Human IgG (H&L).This antibody can be used for detection of the target molecule in samples from Human.
What are the applications and the optimal dilutions of the product Antibody against PARP (Clone 10H)for analysis? - Immunocytochemistry : 5-20 ?g/ml | Western Blot (10 ?g/ml using colorimetric methods | <2 ?g/ml for ECL) | Immunoprecipitation and ELI
Description: Goat IgG Secondary antibody Clone: pAb against UltraPolymer Goat Anti-Human IgG (H&L) HRPfor application in IHC.The immunogen is Human IgG (H&L).This antibody can be used for detection of the target molecule in samples from Human.
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human Goat Anti-TEB4 / MARCH-VI (aa 701 to 712) . This antibody is tested and proven to work in the following applications:
Description: NFkappaB-p65 is a protein encoded by the RELA gene which is approximately 60,2 kDa. NFkappaB-p65 is localised to the nucleus and cytoplasm. It is involved in activated TLR4 signalling, toll-like receptor signalling pathways, cytosolic sensors of pathogen-associated DNA and immune response. NF-kappa-B is a transcription factor involved in several biological processes. It is held in the cytoplasm in an inactive state by specific inhibitors. Upon degradation of the inhibitor, NF-kappa-B moves to the nucleus and activates transcription of specific genes. NFkappaB-p65 is expressed in the nervous system, intestine, bone, lung and skin. Mutations in the RELA gene result in Ependymoma, Reticuloendotheliosis and Retinitis Pigmentosa-50. STJ94473 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of NFkappaB-p65 protein.
Similarly, bax protein expression was significantly and dose-dependently upregulated in mitochondria, but the corresponding cyt c expression was significantly and dose-dependently down-regulated (p<0.05). Raddeanin A is a potential and effective lung cancer chemotherapy drug, which can induce lung cancer cell apoptosis and inhibit proliferation.
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2 Anti Human Kappa Light Chain Polyclonal Antibody,Biotin)
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