Research Paper Volume 15, Issue 22 pp 13558—13578

Figure 5. Upregulation of PTCHD4-AS promoted MSH2-MSH6 dimerization and activated the ATM-p53-p21 pathway. (A) Schematic diagram of truncated MSH2 used in pull-down assays. (B) After transfection of 3xFlag-MSH2 truncation in 293T cells for 48h, RIP was performed with anti-flag antibody. Western blot analyses of pull-down assays with truncated MSH2 fragments. (C) The relative expression level of PTCHD4-AS in the truncated MSH2 RIP was detected by RT-qPCR. IgG was used as a negative control. (D, E) Interaction analysis between MSH2, MSH6 and ATM in SGC7901 and MGC803 cells stably overexpressing EV or PTCHD4-AS. Co-IP experiments were performed with anti-MSH2 antibody, and Western blot was used to detect the expression of specific proteins in the precipitates. (F, G) Western blotting analysis of specific protein levels in SGC7901 and MGC803 cells stably overexpressing EV or PTCHD4-AS after transduction with siCtrl or siMSH2 for 48 h. Data are presented as mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001.