Journal: Molecular Endocrinology
Article Title: Knockdown of PRKAR1A , the Gene Responsible for Carney Complex, Interferes With Differentiation in Osteoblastic Cells
doi: 10.1210/me.2013-1152
Figure Lengend Snippet: Differential expression of PKA regulatory and catalytic subunits in MC3T3-E1 preosteoblasts. A, MC3T3-E1 cells were grown in the presence (+) or absence (−) of osteoblast differentiation medium (ODM). Transcript levels were determined using QPCR, with expression levels normalized to glyceraldehyde-3-phosphate dehydrogenase. Data are expressed as the mean ± SD of three independent experiments. *, P < .05, **, P < .01 compared with undifferentiated cells. B, Protein lysates of MC3T3-E1 cells were analyzed by Western blotting. Actin was used as the internal control. Numbers below the panels indicate normalized (relative to actin) levels of PKA regulatory and catalytic subunits, expressed relative to the value for control shRNA, which is arbitrarily set at 1.0. This experiment was repeated at least twice with similar results, and a representative blot is shown.
Article Snippet: Osteogenic differentiation was induced using osteoblast differentiation medium (ODM) composed of MEM-α with 50 mg/L ascorbic acid and 10 mM β-glycerophosphate (Sigma-Aldrich).
Techniques: Expressing, Western Blot, shRNA