Fig. 2. Yki is phosphorylated by Wts in vivo. (A) Western blot of lysates of wing imaginal discs from wild-type (wt) animals treated with CIP, or untreated wt, ex^e1, ft^G-rv, ex^e1 ft^G-rv and wts^P2, as indicated, run on a conventional 7.5% PAGE gel, probed with anti-Yki and, as a loading control, anti--Actin. (B) Western blot of lysates of wing imaginal discs from wt or wts^P2 animals treated with CIP, or untreated wt, ex^e1, ft^G-rv, ex^e1 ft^G-rv and wts^P2 as indicated, run on a Phos-Tag gel (25 µM Phos-Tag), probed with anti-Yki and, as a loading control, anti-Actin. Arrow indicates band corresponding to unphosphorylated Yki. (C) Conventional 4-15% PAGE gel, probed with anti-Yki (upper panel), and, as a loading control (lower panel), anti--Actin. Lane 1 shows S2 cell lysate treated with ds RNA corresponding to GFP (control). Lane 2 shows lysate from S2 cells treated with ds RNA corresponding to yki. Yki protein (arrow) is reduced, but a faint background band (above) is unaffected.