Figure 3
From: A STRIPAK component Strip regulates neuronal morphogenesis by affecting microtubule stability
Strip physically and genetically interacts with TBCD.
(a,b) Representative images of wild-type (a) or stripdogi (b) DL1 PN clones (green). Bruchpilot (Brp) staining in magenta. Scale bar is 25 μm. Asterisk: cell body. Yellow and white dotted circles indicate the mushroom body (MB) and the lateral horn (LH), respectively. (c) Immunoprecipitation of S2 cell lysates treated with control or TBCD dsRNA for 6 days. Endogenous TBCD was co-immunoprecipitated when endogenous Strip was precipitated by anti-Strip antibody in control dsRNA-treated S2 cells but not in TBCD dsRNA-treated cells. Red triangles indicate endogenous TBCD. (d) Immunoprecipitation of S2 cell lysates expressing c-myc tagged TBCD (TBCD-myc). TBCD-myc was co-immunoprecipitated when endogenous Strip was precipitated by anti-Strip antibody. (e) Immunoprecipitation of S2 lysates expressing Strip. Endogenous TBCD was co-immunoprecipitated when Strip was precipitated by anti-Strip antibody. (f) Representative image of S2 cells expressing TBCD-myc. Magenta: TBCD-myc, green: endogenous Strip. (g,h) Representative images of single-cell clones of DL1 PNs expressing shRNA against TBCD (shRNA-TBCD) in wild type (g) or stripdogi (h) PNs. (i,j) Representative images of single-cell clones of DL1 PNs overexpressing TBCD in wild type (i) or stripdogi (j) PNs. The right panels in (h) and (j) show PNs exhibiting the ‘extremely short axon’ phenotype, i.e., the axon did not exit the antennal lobe (white circle) and did not enter the mushroom body (MB, yellow dotted circle) or the lateral horn (LH, white dotted circle). Scale bar is 25 μm. Genotypes: (a) y w, hs-FLP122, UAS-mCD8GFP/y w (or Y); GH146-Gal4, UAS-mCD8-GFP/+; tubP-Gal80, FRT2A/FRT2A, (b) y w, hs-FLP122, UAS-mCD8GFP/y w (or Y); GH146-Gal4, UAS-mCD8-GFP/+; tubP-Gal80, FRT2A/stripdogi, FRT2A, y+, (g) y w, hs-FLP122, UAS-mCD8GFP/y w (or Y); GH146-Gal4, UAS-mCD8-GFP/UAS-shRNA-TBCD#1; tubP-Gal80, FRT2A/FRT2A, (h) y w, hs-FLP122, UAS-mCD8GFP/y w (or Y); GH146-Gal4, UAS-mCD8-GFP/UAS-shRNA-TBCD#1; tubP-Gal80, FRT2A/stripdogi, FRT2A, y+, (i) y w, hs-FLP122, UAS-mCD8GFP/y w (or Y); GH146-Gal4, UAS-mCD8-GFP/UAS-TBCD; tubP-Gal80, FRT2A/FRT2A, (j) y w, hs-FLP122, UAS-mCD8GFP/y w (or Y); GH146-Gal4, UAS-mCD8-GFP/UAS-TBCD; tubP-Gal80, FRT2A/stripdogi, FRT2A, y+.
