Journal: Biology of reproduction

Article Title: Antiproliferative effects of adiponectin on human trophoblastic cell lines JEG-3 and BeWo.

doi: 10.1095/biolreprod.108.070573

Figure Lengend Snippet: FIG. 2. Phosphorylation of (P-PRKA) by adiponectin in BeWo and JEG-3 cells. Effects of adiponectin on PRKA activation in BeWo (A and C) and JEG-3 (B and D) cells. Cells were maintained in a serum-free culture medium overnight. Cells were then exposed to human recombinant adiponectin (250 ng/ml) or AICAR (500 lg/ml). At the indicated times, cellular extracts were prepared and immunoblotted with anti-phospho-PRKA or with anti-total PRKA. A and B) Western blot analysis from a representative experiment. C and D) Densitometric analysis of PRKA immunoblots. The figure shows means of active phospho-PRKA/total PRKA ratios. Results are means 6 SEM obtained from four to six separate experiments and are expressed as percentages of control value (untreated). *P , 0.05, Wilcoxon test; ns, nonsignificant.

Article Snippet: Proteins were transferred to PVDF membrane and were blocked in buffer A with 2.5% gelatin during 2 h. Membranes were incubated overnight at room temperature in buffer A with 2.5% gelatin rabbit polyclonal anti-phospho-PRKA antibody (1:700 dilution, #2531; Cell Signaling Technology, St-Quentin-en-Yvelines, France) or rabbit polyclonal anti-total PRKA antibody (phosphorylated and nonphosphorylated forms) (1:1000 dilution, #2532; Cell Signaling Technology).

Techniques: Phospho-proteomics, Activation Assay, Recombinant, Western Blot, Control