Figure 4

Ecdysone directly induces glomerular mesangial cell activation and extracellular matrix accumulation. (A) Mouse mesangial cells were treated with ecdysone (10−7 M) or vehicle for 48 hours. Cells were fixed and processed for staining for fluorescent immunocytochemistry staining for α-SMA, a marker of mesangial activation, or the extracellular matrix component fibronectin. Cells were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Bars = 20 µm. (B) Mesangial cell lysates were collected at 48 hours and processed for western blot analysis for α-smooth muscle actin (α-SMA) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). (cropped blots are shown as indicated by the lines and full-length blots are included in the Supplementary Figure 3).