Fig. 6

CircHERC1 binds FOXO1 and sequesters it in the cytoplasm. (a) Western blot analysis of pFOXO1 and FOXO1 in NCI-H3255 cells with circHERC1 overexpression or knockdown. (b) Western blot analysis of pFOXO1 and FOXO1 in the cytoplasm and nucleus of NCI-H3255 cells with circHERC1 overexpression or knockdown. (c) IF assays to detect FOXO1 subcellular localization in A549 cells after alteration of the expression of circHERC1 The scale bars are 15 μm. (d) Western blot analysis of FOXO1 in the cytoplasm and nucleus in NCI-H3255 cells transfected with the control vector, control vector treated with LY2940002, circHERC1 overexpression vector, and circHERC1 overexpression vector treated with LY2940002, or cells pretreated with LY2940002 transfected with circHERC1 overexpression vector. (e) RNA pulldown assays to evaluate the interaction between circHERC1 and pFOXO1 or FOXO1 using biotinylated probes against circHERC1. (f) RIP assays to evaluate the interaction between circHERC1 and FOXO1 after alteration of the expression of circHERC1 and treatment with AS1842856 using a FOXO1 antibody. **p < 0.01. (g) Western blot analysis of pAKT in NCI-H3255 cells with circHERC1 overexpression treatment with AS1842856. (h) Western blot analysis of pFOXO1 and FOXO1 in the cytoplasm and nucleus in NCI-H3255 cells transfected with the control vector, circHERC1 overexpression vector, or FOXO1 knockdown vector and cotransfected with the circHERC1 overexpression vector and FOXO1 knockdown vector