Fig. 3: RUNX1 interacts with NPM1 for promoting the expression of ECM-associated genes.

A Anti-FLAG antibody-pulled down samples from FLAG-RUNX1 expressing N9 cells were analyzed by the SDS-PAGE and silver staining. B, C Anti-RUNX1 or anti-NPM1 antibody co-IPed samples were analyzed by WB. D PLA was performed for detecting the interaction between RUNX1 and NPM1. E Schematic of various RUNX1 and NPM1 truncations. The domains annotated with residue numbers and truncations of RUNX1 and NPM1 were shown. RHD: runt-homology domain. TAD: transactivation domain. OligoD: oligomerization domain. HistonD: histone binding domain. NBD: nuclear acid binding domain. F, G Total cell lysates from HEK 293 T cells expressing different truncations of RUNX1 and NPM1 were IPed and immunoblotted with antibodies against HA or MYC tags. Red asterisk: heavy chain. The RUNX1 H, I The mRNA levels of RUNX1, FN1, COL4A1, and LUM in N9 and TBD0220 cells with NPM1 KD were detected by qRT-PCR assays. GAPDH served as the internal control. J, K The protein levels of RUNX1, FN1, COL4A1, and LUM in N9 and TBD0220 cells with NPM1 KD were detected by WB assays. GAPDH or β-Tubulin served as the internal control. L, M The levels of FN1, COL4A1, and LUM in the culture supernatants of N9 and TBD0220 cells transfected with siNPM1 were measured by ELISA. N ChIP results of the promoters of FN1, COL4A1, and LUM genes enriched by NPM1 in RUNX1 KD N9 cells. Student’s t-test for the two-group analysis, and one-way ANOVA for comparison of multiple groups. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.