GAPDH Antibody - #AF0911

Figure 5. TNFRSF14 enhances adhesion ability of DSC through MMP9. (A) MMP9 levels in vehicle or rapamycin-treated control or siTNFRSF14 DSCs (n = 6 per group) were analyzed by RT-PCR. (B) The transcription level of MMP9 in ESCs (n = 5) and DSCs (n = 6) was analyzed by RT-PCR. (C) The MMP9 transcription levels in ATG5over ESCs (n = 4), control ESCs (n = 4), siATG5 DSCs (n = 8) and control DSCs (n = 8) were analyzed by RT-PCR. (D) The level of MMP9 protein in ATG5over (n = 3) and control ESCs (n = 3) was detected by western blotting. (E) After treatment with different concentrations (0, 1, 10, 100, 1000 µM) of edaravone, the expression of MMP9 in DSCs (n = 6) was evaluated by western blotting. (F) The expression of adhesion molecules in DSCs (n = 6) after 1 mM edaravone treatment was analyzed by flow cytometry. (G) After treatment with C57 pregnant mice by vehicle (1% DMSO) (n = 5) or edaravone (n = 5, 2 mg/kg, daily). The embryo resorption rates of the two groups were compared (H) and the weight of the embryo and placenta was recorded (I) at the gestation of day 13.5. At the gestation of day 7.5, the proportion of PTPRC+ immune cells (J,K) in the uterus, the proportion and number of CD3− KLRB1+ NK cells (J,L) and the expression of adhesion molecules on VIM+ USCs (M,N) of pregnant mice were analyzed by flow cytometry (n = 7 per group). Data were presented as mean ± SEM or median and quartile and analyzed by t test or ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, NS: no significance

Fig. 3 USP7 stabilizes YY1 expression through deubiquitination. (A) Effect of USP7 on YY1 expression in PLC-PRF-5 and HepG2 cells as detected by Western blot. (B) Changes in YY1 expression after transfection with different amounts of Flag-USP7 vector. (C) Western blot analysis of the change in YY1 expression in siUSP7 PLC-PRF-5 and HepG2 cells treated with or without MG132. (D) Effect of USP7 on YY1 degradation in PLC-PRF-5 cells transfected with siUSP7 or Flag-USP7 vector and treated with CHX. (E) Effect of USP7 on YY1 ubiquitination in PLC-PRF-5 cells treated with siUSP7 and Flag-YY1. Flag beads were used for immunoprecipitation. (F) Effect of USP7 on YY1 ubiquitination in PLC-PRF-5 cells treated with Flag-USP7. YY1 antibody was used for immunoprecipitation. Data are expressed as the mean ± SD (*P 

Figure 1. The decidualization process is accompanied with senescent DSCs. (A) Subpopulation of stromal cells of secretory endometrium and decidua in t-SNE plots through single-cell sequencing. (B) QuSAGE analysis of ESCs and DSCs through single-cell sequencing. (C) Bubble diagram presented the average expression of cell cycle and senescence-associated secretory phenotype (SASP) related genes based on the results of single-cell sequencing. (D) Human ESC cell line (hESCs) was treated with 8-bromo-cAMP (0.5 mM) plus MPA (1 μM) for different times, and western blotting assay was used to detect the expression of CDKN2A, CDKN1A, TP53 (indicator of cell senescence) and PRL. (E) SAβG staining (indicator of cell senescence) and statistical data of decidualized hESCs (n = 3 biological replicates per group). (F) Primary ESCs were treated with 8-bromo-cAMP (0.5 mM) plus MPA (1 μM) for 4 days, and the protein of CDKN2A, CDKN1A, TP53, and PRL were measured by western blotting (n = 3 biological replicates per group), relative expression levels of proteins were standardized using internal reference GAPDH. (G) SAβG staining in primary ESCs (n = 4 biological replicates) and primary DSCs (n = 4 biological replicates). Statistical data were presented as mean ± SEM. **P 

Fig. 3 β2AR inhibits cAMP production in CIA-FLSs by coupling with Gαi instead of Gαs. A Intracellular cAMP production in ISO (1 μM)-treated normal rat FLSs that were pretreated with Bar (10 μM), Gαs siRNA, or Gαi siRNA was detected in the FRET system. B The membrane and cytosolic distribution of β2AR after ISO stimulation was evaluated in normal and βarr2-deficient rat FLSs. C The cytosolic expression of β2AR was quantified. D The membrane expression of β2AR was quantified. E–G The binding of β2AR with Gαs or Gαi in FLSs from normal and CIA rats was determined by co-IP, and the data were analysed. H-J The expression of Gαs, Gαi, and β2AR in normal and CIA rat FLSs was analysed using input samples. K The effect of knocking down βarr2, Gαs, or Gαi on ISO-induced FLS viability was evaluated by a CCK-8 assay. L The effect of knocking down βarr2, Gαs, or Gαi on ISO-induced FLS migration was analysed. M The effect of knocking down βarr2, Gαs, or Gαi on ISO-induced FLS invasion was analysed. The data are presented as the means ± SEMs; *p 

Fig. 1: LAP2α interacted with shelterin complex and LAP2α deficiency induced dysfunctional telomeres of ALT-positive cells. A, B PLA of LAP2α and subunit of shelterin complex in U2OS cells and Hela cells transfected with control or LAP2α siRNA. Red dots represent PLA signals. C U2OS and Hela cells were infected with control or LAP2α siRNA for 3 days. Cell lysates were subjected to western blot analysis with anti-LAP2α and anti-GAPDH antibodies. GAPDH was used as the loading control. D Quantification of the LAP2α relative protein level in comparison to GAPDH and to siNC was calculated by ImageJ software. Error bars represent the mean ± SEM of four independent experiments. Two-tailed unpaired Student’s t-test was used to calculate p-values. **p 

Fig. 4 10− 6 M insulin inhibits the gene and protein expressions of the IIS-related receptors and substrates in human DPSCs. A 10− 6 M insulin down-regulated the mRNA levels of INSR, IGF1R, and IRS1 in DPSCs at day 3 and day 7. B 10− 6 M insulin inhibited the protein expressions of INSR, IGF1R, and IRS1 in DPSCs at day 7. Representative western blotting (left) and quantification analysis (right). Full-length blots/gels are presented in Supplementary Fig. 2. Data are expressed as the mean ± SD of n = 3.

Fig. 4 Effects of dietary xylanase on the inflammation response in Nile tilapia. Gene level of A Nuclear factor kappa B (nfkb), B Interleukin 1 beta (il1β) and C Caspase3 (casp3) (n = 6), D Western blot analysis of intestinal P65 and IL1β (n = 3), E Quantitation of the levels of P65 and IL1β normalized to that of GAPDH (n = 3). Data was expressed as mean ± SEM. SM, fish fed with soybean diet; SMC, fish fed with soybean diet supplemented with 3,000 U/kg xylanase. The significant differences between two group were presented at P