Figure 5

Regulation of miR-99a expression in different culture conditions. (A) The levels of miR-99a expression in OEC-M1, TW2.6, and HSC3 oral cancer cells treated with 5 μM 5-Aza-dC for 96 hours were determined using qRT-PCR analysis. Expression of miR-99a was normalized against an endogenous control U6. Data from 5-Aza-dC-treated OSCC cell lines were normalized to those from untreated controls. (B) Immunoblot analysis revealed reduced levels of total and phosphorylated IGF1R protein in non-silencing microRNA expressing controls (OEC-M1 NS) with serum starvation. Ratios were determined by dividing the protein levels in serum-starved cells with that in complete medium (CM). The means in the lower panel were measured by averaging the ratios from independent blots. Bar, SE; *p < 0.05. (C) MiR-99a expression increased in serum starvation in a time-dependent manner. Expression of miR-99a was normalized against an endogenous control U6. The relative expression of miR-99a was determined by normalizing the expression of miR-99a in OEC-M1 NS cells in serum starvation to that in complete medium (CM). Bar, SE; **p < 0.01.