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The following primary antibodies were used at indicated dilution: rabbit monoclonal to phospho-RIPK1 (Ser166, clone D1L3S) diluted 1:1 000 (65746, Cell Signaling Technology, Danvers, MA, USA); rabbit polyclonal to PARP1 diluted 1:1 000 (ab137653, Abcam); mouse monoclonal to SQSTM1/p62...
necrosulfonamide (NSA), trolox (Vit E), and U0126 were purchased from Sigma-Aldrich. Captopril (CAP) was purchased from Cell Signaling Technology (Danvers, MA, USA). Z-VAD-FMK was purchased from Enzo Life Sciences
The primary antibodies against Sp1, MMP-9, Cyclin D1 were purchased from Cell Signaling Technology (Danvers, MA, USA). Protein levels were normalized to total GAPDH or β-actin (Bioworld Technology, MN, USA). Fold-changes were determined. 2.5. Immunofluorescence assay Immunofluorescence of ...
A549 cells were treated with the GSK-3 inhibitor SB216763 (Cell Signaling Technology, Danvers, MA, USA) for 24 h, and H460 cells were treated with rapamycin (KeyGEN Biotech, Nanjing, China) and 3-methyladenine (3-MA, KeyGEN Biotech) for 24 h. Cells were then irradiated with 2-Gy X-...
The immune complexes were detected by enhanced chemiluminescence (ECL; Cell Signaling Technology, Danvers, MA, USA). The integrated density of the band was quantified by Quantity One software (Bio-Rad, Hercules, CA, USA). Statistical Analysis Continuous variables were expressed as the mean ± SD...
Cells were harvested and lysed in protein sample buffer, separated by SDS-PAGE, transferred to nitrocellulose membranes, and immunoblotted using a primary antibody targeted against phospho-ERK or total ERK (Cell Signaling Technology, Danvers, MA) and peroxidase-conjugated secondary antibodies (Bethyl ...
The presence and migration of injected cells in situ were analyzed by IHC using human nuclear specific antibody (HuNu; Cell Signaling Technology, Danvers, MA, USA). Myogenic conversion of injected hAFSCs in vivo was confirmed with real-time PCR using human primers for myogenic lineage markers. ...
The following antibodies were used in this study: HMGCR (ab174830, Abcam, Shanghai, China, 1:2500); TC10 (AB32079, Abcam, 1:2000); FNTB (3283-1, epitomics, 1:10,000); cleaved caspase-3 (9664 s, Cell Signaling Technology, Danvers, MA, USA, 1:1000); Caspase-3 (9662, Cell ...
Immunohistochemistry analysis was conducted as described earlier [26]. Tissue sections were incubated with rabbit monoclonal antibodies against P130cas and YAP (1: 50; Cell Signaling Technology, Danvers, MA, USA). P130cas and YAP staining intensity were scored as 0, 1, 2, and 3, corresponding...