Western Blot Analysis of Kidney Proteins

Mouse kidneys were homogenized in SDS Laemmli buffer and loaded onto 10% SDS-polyacrylamide agarose electrophoresis gels essentially as described previously [27 (link), 47 (link)]. Primary antibodies pERK1/2 (9101 S), pAKT (4060 S), pSTAT3 (9145 S), STAT3 (9139 S), cyclinD1 (2978 S), S6 (2317 S), pS6 (4858 S), PDGFRβ (3169), VEGFR2 (9698), and FGFR1 (9740) from Cell signaling (Danvers, MA, USA); YAP (SC-101199), GAPDH (SC-32233), ERK1/2 (SC-94), cMyc (SC-40) from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA), and α-SMA (ab5694; Abcam, Cambridge, MA, USA); and secondary antibodies, anti-mouse (P0447) and anti-rabbit (P0448) from Dako (Santa Clara, CA, USA) and ECL reagent from Amersham (GE Health care, Buckinghamshire, UK) were used.