Fig. 5

HDAC1 is necessary for SOX4 promoting colorectal cancer stemness. a HDAC1 is necessary for SOX4 promoting sphere-forming capacity. Indicated cells were seeded in ultra-low attachment 6 well plate. The number of spheres was counted after 10 days. b HDAC1 depletion abolished the effect of SOX4 on the self-renewal capacity of colorectal cancer cells on serial passage. The sphere number of indicated primary, secondary, and tertiary passaged cells was counted after 10 days. c The depletion of HDAC1 abolished the effect of SOX4 on the sphere-forming cell frequency of colorectal cancer cells. Indicated cells were maintained in 96-well U-bottomed culture plates at a density of 10, 5 or 1 cells per well and cultured for 10 days. The sphere-forming cell frequency was calculated by ELDA software. d Knockdown of HDAC1 attenuated the effect of SOX4 on tumor-initiating cell frequency of colorectal cancer cells. Indicated cells were injected into the subcutaneous tissues of 6-week-old nude mice at a density of 21, 7, 3 cells per mouse. The number of mice developed tumors was counted after 35 days. The tumor-forming cell frequency was calculated by ELDA software. e Knockdown of HDAC1 attenuated the effect of SOX4 on the expression of cancer stem cell markers of colorectal cancer cells. The surface protein levels of CD44 and CD133 in indicated HCT-116 cells were analyzed by flow cytometry. Data are represented as mean ± s.d.; *P<0.05, **P<0.01, ***P<0.001; two-tailed Student’s t-test