Fig. 1

Highly expressed FBXO22 is required for the growth of AML especially MLLr AML cells. A FBXO22 mRNA expression level was analyzed in GSE13159 database. B FBXO22 protein in BM mononuclear cells (BMMC) of AML (including 1 case of M1 or M2, 9 cases of M4 and 4 cases of M5 AML samples) and non-leukemic individuals was analyzed by Western blot with Histone 3 (H3) as loading control. C, D AML 3# MNCs were infected with shFBXO22#1, shFBXO22#2 and shNC, and FBXO22 protein was immunoblotted with β-actin as loading control (C). Cell numbers were counted at the indicated days (D). E Colony-forming assay (left) for AML 3# MNCs infected with shNC, shFBXO22#1 and shFBXO22#2, and colony numbers at day 10 were counted (right, n = 3). F, G THP-1 cells were infected with shFBXO22#1, shFBXO22#2 and shNC, and FBXO22 protein was immunoblotted with β-actin as loading control (F), and cell proliferation was examined by CCK-8 assay at the indicated days (G). H Colony-forming assay (left) for THP-1 cells infected with shNC, shFBXO22#1 and shFBXO22#2, and colony numbers at day 7 were counted (right, n = 3). I–K Cell-cycle (I) or apoptosis (J) analysis for THP-1 cells infected with shNC, shFBXO22#1 and shFBXO22#2 (n = 3). Indicated proteins were immunoblotted with α-tubulin as loading control (K). L, M THP-1 cells infected with shNC or shFBXO22#1 were stably transfected with EV or Flag-tagged shRNA-resistant FBXO22, and indicated proteins were immunoblotted with β-actin as loading control (L), and cell proliferation was examined by CCK-8 assay at the indicated days (M). N Colony-forming assay (left) for the indicated THP-1 cells, and colony numbers at day 7 after plating were counted (right, n = 3). O Apoptosis analysis for indicated THP-1 cells (n = 3). Error bars denote mean ± SD. Statistical significance was determined by two-tailed unpaired t test (A, E, H–J and N–O) or two-way ANOVA (D, G and M), and the P values were shown. All experiments were repeated three times with similar results