Figure 4.
Figure 4. Bnl is cleaved in the Golgi network of Bnl-producing cells. (A and B) Projection images of lower Z-stacks of the disc bnl source showing detergent-free αGFP immunostaining (αGFPex; red) when bnl-Gal4 expressed Bnl:GFP1 (A) and Bnl:GFP3 (B). White, phalloidin-Alexa Fluor 647 to mark actin-rich cell outlines; arrowhead, intracellular Bnl:GFP (only green); arrow, surface-localized Bnl:GFP (green+red); dashed line, ASP in the upper Z-stacks (not shown). (C and C′) αGM130-stained (red) optical sections of wing disc bnl-source expressing Bnl:GFP1. (D–J′) Single optical sections of αHA-immunostained (red) disc bnl-source (D–G′) and S2 cells (H–J′) expressing either Bnl:HA1GFP3 or M1 and marked with α-Stx-16 or αGM130 (blue) as indicated. Arrow, truncated Bnl:GFP3 derivative; arrowhead, uncleaved Bnl:HA1GFP3 or M1 mutant; merged (D–J) and split (D′–J′) blue channels shown. Scale bars: 20 µm (A–C); 5 µm (C′–G′); 10 µm (H–J′).

Bnl is cleaved in the Golgi network of Bnl-producing cells. (A and B) Projection images of lower Z-stacks of the disc bnl source showing detergent-free αGFP immunostaining (αGFPex; red) when bnl-Gal4 expressed Bnl:GFP1 (A) and Bnl:GFP3 (B). White, phalloidin-Alexa Fluor 647 to mark actin-rich cell outlines; arrowhead, intracellular Bnl:GFP (only green); arrow, surface-localized Bnl:GFP (green+red); dashed line, ASP in the upper Z-stacks (not shown). (C and C′) αGM130-stained (red) optical sections of wing disc bnl-source expressing Bnl:GFP1. (D–J′) Single optical sections of αHA-immunostained (red) disc bnl-source (D–G′) and S2 cells (H–J′) expressing either Bnl:HA1GFP3 or M1 and marked with α-Stx-16 or αGM130 (blue) as indicated. Arrow, truncated Bnl:GFP3 derivative; arrowhead, uncleaved Bnl:HA1GFP3 or M1 mutant; merged (D–J) and split (D′–J′) blue channels shown. Scale bars: 20 µm (A–C); 5 µm (C′–G′); 10 µm (H–J′).

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