Fig. 4

CD274 interacts with JNK to increase Cyclin D2 level. a The phospho-JNK and total JNK levels in WT and CD274-null LICs were determined by Western blotting analysis. b The phospho-JNK and total JNK levels were evaluated in CD274-over-exprssed (OE) 293T cells by Western blotting analysis. c Strep II tagged CD274 was overexpressed in 293T cells, followed by immunoprecipitation with Strep II beads and Western blotting analysis for the levels of CD274, phospho-JNK, and JNK. d Fc-tagged JNK was overexpressed in 293T cells, followed by the immunoprecipitation with protein A/G beads and Western blotting analysis for the level of CD274 and JNK. e The Cyclin D2 level was evaluated in JNK-overexpressed (OE) 293T cells by Western blotting analysis. f The knockdown efficiency of shRNAs (#1-#3) targeting JNK was evaluated in 293 cells by Western blotting analysis. g WT and CD274-null LICs were knocked down with shRNA#1 and cultured in solution medium for 6 days in vitro. h Representative images for the colony forming units of WT and CD274-null LICs infected with shRNA#1 targeting JNK. i, j Colony number (i) and total cell number of colonies (j) in h were calculated (n = 3, ***P < 0.001, **P < 0.01). k Working model for the function of CD274 in leukemogenesis