Figure 2. Drosophila Hh is secreted in Lpp-associated and Lpp-free forms.

(A) Hh levels in hemolymph and whole extracts of wild-type and hh^TS larvae at restrictive temperature, analyzed by WB. Hemolymph loading control is a secreted GFP expressed from the tubulin promoter. (B) Hemolymph Hh levels in wild-type and disp mutant larvae, analyzed by WB. Loading control is Cv-d. (C) Density of Hh in hemolymph of wild-type and Lpp RNAi larvae, analyzed by Optiprep density gradient centrifugation and WB. Equal amounts of hemolymph (normalized by protein) were analyzed. (D) Hemolymph Hh levels in larvae overexpressing Hh in imaginal discs with en105-GAL4, analyzed by WB. Loading control is Cv-d. (E) Density of hemolymph Hh in larvae overexpressing Hh in imaginal discs, analyzed by Optiprep density gradient centrifugation and WB. (F) Density of Hh lipid modification mutants (Hh^C85S, Hh-N, Hh-N^C85S), secreted into the hemolymph from the fat body (FB) with lpp-GAL4, analyzed by Optiprep density gradient centrifugation and WB.