Fig. 7

The roles of PAX1 mutants associated with SCID in regulation of Wnt signaling pathway. A Sanger sequencing results of wild type (WT) control and SCID associated PAX1 mutation constructs. B-C TopFlash luciferase reporter assays performed in HEK293FT cells transfected with empty vector control, WT or PAX1 mutation constructs without (B) or with (C) treatment of CHIR99021 (CHIR). * represents significant decrease compared with control, while # represents significant increase compared with PAX1 WT. D HEK293FT cells were transfected with HA-tagged TCF7L2 and empty vector control, PAX1 WT or PAX1 mutation construct (flag-tagged). TCF7L2 levels were analyzed by western blotting with anti-HA antibody. GAPDH was used as loading control. E A series of PAX1 mutation constructs with flag-tag were co-transfected with HA-tagged TCF7L2 in HEK293FT cells. Co-IP and western blotting assays were performed to test the interactions between TCF7L2 and PAX1 proteins. F Interactions between TCF7L2 and PIASy were assessed by co-IP and western blotting with empty vector control, PAX1 WT or PAX1 mutation constructs cotransfection in HEK293FT cells. Co-IP was done with anti-HA antibody. TCL and IP samples were assayed by western blotting using anti-myc antibody for PIASy and anti-flag antibody for PAX1 proteins, respectively. GAPDH was used as loading control