Up-regulation of CDKN1A (p21Waf1/Cip1) was observed in INO80 complex knockdown HeLa or 293T cells.
(A) Verification of the mRNA of select genes from gene expression profiles. HeLa cells were transfected with 15pmol INO80- or hArp8-siRNA and siNT (as control). 48 hours after transfection, cells were harvested, and select genes from gene expression profiles were assessed by RT-qPCR. Bar graphs show ratios of RT-qPCR signals to GAPDH (all signals normalized to siNT). Error bars represent the standard error of the mean of 3 independent experiments. (B) Elevation of p21 protein expression in INO80- or hArp8-siRNA knockdown 293T cells. Cells were treated with indicated siRNA. 48 hours later, proteins were detected with WB using indicated antibodies. (C) High expression of p21 protein in cells as assessed by immunofluorescence. INO80- or hArp8-siRNA (15 or 30pmol) and siNT control was transfected into HeLa cells. 48 hours after transfection, immunofluorescence staining was performed. DAPI staining shows total nuclei. p21-positive cells were counted, and the percentage in the total cell numbers was represented in bar graph (D). Error bars represent the standard error of the mean of 2 independent experiments. **p < 0.01 in comparison with siNT control (Student t test).
