Fig. 4
The effect of IMPDH2 on EMT and cell-cycle transition of CRC cells. (a) The spindle cell phenotype of IMPDH2-overexpressed cells (SW480/IMPDH2 and LoVo/IMPDH2) and the epithelial phenotype of CRC cells from its control group (Vector) showing epithelial-to-mesenchymal transition (EMT) induced by IMPDH2 overexpression. (b) IMPDH2 overexpression induces hallmarks of the EMT, including loss of E-cadherin and accumulation of Vimentin and Snail in CRC cells. (c and d) Immunofluorescent staining of E-cadherin and Fibronectin in SW480/IMPDH2 and LoVo/IMPDH2 Cells. (e) GSEA showing a significant association between IMPDH2 expression and CELL_CYCLE and PI3K_AKT_MTOR_SIGNALING signaling pathway. The top portion of the figure plots the enrichment scores for each gene, whereas the bottom portion of the plot shows the value of the ranking metric moving down the list of ranked genes. Y-axis: value of the ranking metric; X-axis: the rank for all genes. NES, normalized enrichment score. (f) Cells proportion in various phases of the cell cycle. Cells were stained with Propidium Iodide (PI) and analyzed by flow cytometry. Mean ± SD (n = 3). (g) Western blotting analysis of p21Cip1, p27Kip1, cyclin D1, and Ki-67 proteins in IMPDH2-overexpressed cells or IMPDH2 shRNA-infected cells. GAPDH was used as a loading control. *P < 0.05; **P < 0.01. (h) Real-time qPCR analysis of p21Cip1, p27Kip1, Ki-67 and cyclin D1 mRNA expression in IMPDH2-overexpressed cells (upper panel) or IMPDH2 shRNA-infected cells (lower panel). Expression levels were normalized to GAPDH. Mean ± SD (n = 3)