Fig. 3. Western immunoblot analysis of Hts proteins in ovary and S2 cell extracts. (A,B) Ovary extracts from virgin w¹¹¹⁸ flies blotted with htsF and htsRC. (A) Both the Add doublet band at 90 kDa and the 80 kDa Ovhts-Fus band can be seen. (B) Only the Ovhts-RC doublet is detected. (C) Ovary extract from flies expressing tagged hts transgenes blotted with anti-GFP antibody. A 75 kDa nonspecific band is present in all lanes. The ShAdd::Ven band is of the predicted size. The bands in Cer::Ovhts-Fus (110 kDa, ^*) and Ovhts-RC::GFP (90 kDa, ^**) are of the sizes predicted for the respective cleavage products fused to the fluorescent proteins. (D,E) Protein extracts from S2 cells expressing Ovhts blotted with 1B1 and htsRC. (D) S2 cells express endogenous Add proteins that can be seen at 90 kDa. Bands representing both full-length Ovhts protein and Ovhts-Fus are indicated. (E) Bands representing both full-length Ovhts protein and Ovhts-RC are indicated. All Hts proteins consistently ran 10-20 kDa larger than predicted.