Fig. 3. The difference in S182N and Cos2 activities is not due to differences in expressed protein levels. Western blot showing overexpression of mutant and wild-type cos2 results in production of comparable levels of each protein in imaginal wing disc lysates (30 µg protein loaded per lane). Arrows indicate migration differences between GFP-tagged proteins and their untagged counterparts. The overexpressed protein is indicated above each lane on the blot. WT, wild-type lysates; Cos2, Hh indicates overexpression of both Cos2 and Hh; S182N-GFP, Hh indicates over-expression of both Hh and S182N-GFP. All experiments were performed using independently isolated fly lines of tagged and untagged Cos2 proteins with the same results in all experiments. 71B Gal4 was the driver for all constructs. BAP111 is a ubiquitous nuclear protein used as a loading control.