Fig. 4

In vitro expression experiments in HEK293T cells show decreased MSH4 expression with c.2222-2225delAAGA (MUT1) variant, but no change with c.2347 A > G (MUT2) variant. (A) Sanger sequencing confirmed the successful construction of MSH4 expression plasmids. (B) RT-qPCR analysis showed MSH4 mRNA downregulation in total RNA extracted from HEK293T transfected with MUT1 expression plasmids. Results were normalized against GAPDH. Data was presented as mean ± SEM. **P = 0.01. (C) Western blot analysis of protein extracts from HEK293 cells transfected with WT, MUT1 and MUT2 expression plasmids. MSH4 was tagged with a Flag epitope and detected with a Flag antibody, while GAPDH served as a loading control. Data was presented as mean ± SEM. *P = 0.05