Figure 2
Lnk acts upstream of Chico ensuring its proper localisation. (A-D) Low phospho-PKB levels in chico−/− MARCM clones are not rescued by overexpression of lnk-CFP. Clones are marked by GFP expression (’ panels) and outlined by a dashed line in the detail pictures (”” panels). Phospho-PKB staining is shown in ” panels. (A-A””) control clones. (B-B””) Levels of phospho-PKB (B”) are decreased in chico−/− clones. (C-C””) Overexpression of lnk-CFP within control clones promotes high levels of phospho-PKB (C”). (D-D””) lnk-CFP overexpression within chico−/− clones does not rescue low levels of phospho-PKB (D”). (E-G) Chico overexpression restores localisation of the tGPH reporter at the membrane in lnk−/− salivary glands. (E) tGPH reporter localises at the membrane in wild-type salivary glands. (F) lnk4Q3 salivary glands show diffuse tGPH reporter signal in the cytoplasm. (G) tGPH reporter is recovered to the membrane of lnk4Q3 tissue by overexpressing chico. (H-K) Subcellular localisation of RFP-tagged Chico. The RFP signal is shown in ’ panels. Actin staining marks the cellular cortex in ” panels. Merged pictures are shown in ”’ panels. Signal intensities along the lines indicated in the ”’ panels are displayed in ”” panels. Quantifications in Additional file 1: Figure S1A. (H-H””) Cortical localisation of Chico-RFP in a wild-type situation. (I-I””) Chico-RFP in a lnk mutant background. (J-J””) A PH domain mutant form of Chico localises at the cortical membrane. (K-K””) Chico-PH*-RFP in a lnk mutant background. Number of samples analysed: (A) n=9, (B) n=9, (C) n = 10, (D) n = 12, (E) n=5, (F) n=5, (G) n=6, (H) n=6, (I) n=9, (J) n=6, (K) n = 6. Scale bars represent 50 μm.