Fig. 5.

Steady-state mRNA and protein levels modulation of IGF upon incubation of C₂C₁₂ cells with 1,25-D3. Cultures of C₂C₁₂ cells were incubated as in Fig. 1 for 4 d. Total RNA and whole-protein extracts were isolated for qRT-PCR and Western blottings, respectively. A, Mean ± sem corresponds to experiments done in triplicate for IGF-I (left panel); **, P < 0.01 and IGF-II (right panel); **, P < 0.01. B, Western blottings with the respective densitometric analysis for IGF-I; ***, P < 0.001. C, Western blottings with the respective densitometric analysis for IGF-II; ***, P < 0.001. VD1 and VD2 correspond to different pools of two samples each. In both cases, real-time PCR and Western blottings, samples and controls were normalized with GAPDH housekeeping gene. VD, Vitamin treated cells whole extracts.