Full blots for DEVELOPMENTAL-CELL-D-24-00333R2
Description
Figure 7B Western blot analysis of Nr2f2 over-expression in lung c-Kit+ cells after AAV infection. Figure S4B Western blot analysis showing the effects for inducing DTR expression in lung c-Kit+ cells by tamoxifen injection. Figure S5F Western blot analysis of Scf in the lungs of control or SuHx mice. n = 6 in the control group and n = 7 in the SuHx group. Figure S7B Western blot analysis of Nr2f2 down-regulation in lung c-Kit+ cells after AAV infection.
Files
Steps to reproduce
Mouse lung tissues and cells isolated by MACS were lysed in RIPA buffer (P0013B, Beyotime) supplemented with PMSF (P0100, Solarbio, 1:100) and protease/phosphatase cocktail (58725, CST, 1:100). Samples were separated in 8% and 12% SDS-PAGE and transferred to PVDF membranes (IPVH00010, Merck Millipore). After being blocked in 5% skim milk (232100, BD Biosciences) at RT for 1 h, PVDF membranes were incubated in primary antibodies at 4°C overnight and then treated with HRP-conjugated secondary antibodies at RT for 1 h. Membranes were finally visualized by ECL Detection Reagent (36208ES76, YEASEN) and detected on a chemiluminescence image analysis system (Tanon-5200, Tanon). Primary antibodies used in western blot analysis and their dilution ratio were as listed: anti-c-Kit (3074S, CST, 1:1000), anti-Nr2f2 (6434S, CST, 1:1000), anti-DTR (ab66792, Abcam, 1:1000), anti-Scf (ab64677, abcam, 1:1000), anti-β-Actin (AC026, Abclonal, 1:5000).