Figure 4

Non-correlation between hMOF and CA9 is found in renal cell carcinoma cells. A. hMOF protein expression was correlated with acetylation of H4K16 in RCC cell 786–0 and OSRC-2. 293T, 786–0 or OSRC-2 cells were cultured in 6-well tissue culture plates (~2x10⁵ cells/well) in DMEM medium containing 10% fetal bovine serum. Whole cell extracts were subjected to immunoblotting using indicated antibodies (right panel). 293T, 786–0 or OSRC-2 cells from 1 well of a 6 well plate were lysed and total RNA was isolated using Trizol. hMOF and CA9 gene expressions were measured by RT-PCR (left panel) and qRT-PCR (B). C. Effect of hMOF on CA9 mRNA expression levels in RCC cells. RCC 786–0 cells were cultured in 6-well tissue culture plates (~2x10⁵ cells/well) in DMEM medium containing 10% fetal bovine serum. The cells were transfected with 0.25, 0.5, 1 and 2 μg of hMOF cDNAs. 48 hours after transfection, cells were lysed and total RNA was isolated using Trizol. Indicated gene expressions were analyzed by qRT-PCR. D. Effect of hMOF on CA9 protein expression in RCC cells. RCC 786–0 cells were transfected with 0.25, 0.5, 1 and 2 μg of hMOF cDNAs. 48 hours after transfection, cells were harvested and lysed in RIPA buffer. Aliquots of whole cell extracts were subjected to 12% SDS-PAGE, and specific proteins were detected by indicated antibodies.