Research Paper Volume 15, Issue 22 pp 13558—13578

Figure 3. PTCHD4-AS interacted with MSH2-MSH6 dimer in GC. (A) SDS-PAGE of pull-down assay of whole cell lysates from SGC7901 cells using a biotin-labeled control/PTCHD4-AS probe showing PTCHD4-AS binding proteins. (B) Protein characterization of highly unique peptides by mass spectrometry. (C) Western blot analysis of the biotin-labeled probe pull-down eluate in (A). (D) RIP assay of anti-MSH2 or anti-MSH6 antibodies in SGC7901 and MGC803 cells confirmed that PTCHD4-AS interacted with the MSH2-MSH6 dimer. The levels of PTCHD4-AS in the precipitates were detected by RT-qPCR, and IgG was used as a negative control. (E) MSH2 and MSH6 protein levels in the precipitates were detected by Western blotting, and GAPDH was used as a loading control. (F) Representative confocal images and co-localization analysis of PTCHD4-AS (red) and MSH2 (green) in SGC7901 and MGC7901 cells. Data are presented as mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001.