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. Author manuscript; available in PMC: 2015 Jul 3.
Published in final edited form as: Cell. 2014 Jul 3;158(1):143–156. doi: 10.1016/j.cell.2014.05.035

Figure 3. Cytoskeletal tension modulates Yki activity.

Figure 3

A-H) Third instar wing imaginal discs expressing en-Gal4 UAS-dcr2 UAS-GFP or UAS-RFP (green) and A) control B-D) UAS-rok-RNAi, E) UAS-rok.CAT, G) UAS-rok.CAT-KG, H) UAS-sqh.EE, I) UAS-sqh.EE UAS-rok-RNAi, stained for expression of ex-lacZ (magenta), Diap1 (red), DNA (blue/white) or Yki (red/white), as indicated. Lower panels for B, D, and G show vertical sections through the same discs. Dashed yellow line marks A-P compartment boundary. Panels marked prime show individual stains of discs to the left. I) Quantitation of ex-lacZ expression in discs of the genotypes depicted here in panels A, B, E, and G, represented as the mean intensity of nuclear ß-gal staining in P compartment regions divided by mean intensity of nuclear ß-gal staining in A compartment regions; mean DNA staining intensity per nuclear volume is also shown for comparison (N=22 (wild type), 22 (rok RNAi), 10 (ROCK.CAT), 30 (Sqh.EE)). J) Quantitation of Yki in en-Gal4 UAS-dcr2 UAS-rok-RNAi UAS-GFP discs, represented as the mean intensity of nuclear Yki staining in P compartment regions divided by mean intensity of nuclear Yki staining in A compartment regions; mean DNA staining intensity per nuclear volume is also shown for comparison (N=22 (wild type), 19 (rok RNAi)). K,L) Representative western blots and results of quantitation of four independent blots performed on wing disc lysates (20 discs per lane) from nub-Gal4 UAS-dcr2 (labeled as control) and (K) nub-Gal4 UAS-dcr2 with UAS-rok-RNAi (labeled as rok RNAi) or UAS-sqh.EE (labeled as Sqh.EE) or (L) nub-Gal4 UAS-dcr2 UAS-rok.CAT (labeled as ROCK.CAT). The phospho-Yki (pYki) to Yki ratio was normalized to the average ratio in wild-type lysates, using ß-tubulin (Tub) as a loading control. See also Fig. S2.