Research Paper Volume 15, Issue 19 pp 10746—10766

Figure 5. HOXD3 overexpression inhibits proliferation in ccRCC cells. (A, B) The expression level of HOXD3 at mRNA and protein levels was assessed by qRT-PCR and western blotting when 786-O and CAKI-1 cells were transfected with HOXD3-Ctrl and HOXD3. (C) The cell viability of ccRCC cells was examined by MTT assay at 24 h, 48 h, and 72 h. (D) Clone formation assay was used to test the colonies number of 786-O and CAKI-1 cells which were transfected with HOXD3-Ctrl and HOXD3. (E) Cell cycles were determined in 786-0 and CAKI-1 cells after transfection of the HOXD3 overexpression vector or HOXD3-Ctrl. The histogram indicates the percentage of cells at G0/G1, S, and G2/M cell-cycle phases. (F, G) The wound healing and transwell assays were applied for verifying the migration and invasion ability of HOXD3 and control-transfected human ccRCC. Scale bar, 100 μm. (H) The expression of N-cadherin, E-Cadherin, MMP2, CDK4, and CyclinD1 was demonstrated by western blotting in both 786-O and CAKI-1 cells of HOXD3 overexpression (p < 0.01).