Details on the generation of new rhea (talin) and Vinculin alleles can be found in Supplemental Experimental Procedures .
For wing blister quantification, mitotic clones were generated in the wings of heterozygous flies by crossing rhea mutant males to w; P{w[+], Gal4}Vg[BE] P{w[+], UAS::FLP}; P{FRT}2A (with the white+ excised from P{FRT2Aw[hs]}) females. Embryonic phenotype quantification was performed on mutant embryos lacking both maternal and zygotic wild-type talin and/or vinculin, as they were obtained from germline clones generated in heterozygous mutant females by crossing rhea mutant females (with wild-type Vinculin or ΔVinc) to P{hs::FLP}1, y[1] w[118]; P{ovoD1-18}3L P{FRTw[hs]}2A (for genotypes with wild-type Vinculin) or ΔVinc w[-]; P{hs::FLP}38/CyO; P{ovoD1-18}3L P{FRTw[hs]}2A (for genotypes with ΔVinc) males. Heat shocks were performed two times for 1 hr and 15 min each at 37°C at L1 and L2 larval stages. TalinIBS2-mCherry [31 (link)] was kindly provided by H.J. Bellen. The myosin heavy chain mutant used was Mhc[1] [45 (link)], kindly provided by S.I. Bernstein. IBS2-GFP recruitment to muscle attachment sites was performed with UAS::IBS2-GFP [15 (link)] expressed in muscles with P{Gal4-Mef2.R}3 (Bloomington Drosophila Stock Center).
For wing blister quantification, mitotic clones were generated in the wings of heterozygous flies by crossing rhea mutant males to w; P{w[+], Gal4}Vg[BE] P{w[+], UAS::FLP}; P{FRT}2A (with the white+ excised from P{FRT2Aw[hs]}) females. Embryonic phenotype quantification was performed on mutant embryos lacking both maternal and zygotic wild-type talin and/or vinculin, as they were obtained from germline clones generated in heterozygous mutant females by crossing rhea mutant females (with wild-type Vinculin or ΔVinc) to P{hs::FLP}1, y[1] w[118]; P{ovoD1-18}3L P{FRTw[hs]}2A (for genotypes with wild-type Vinculin) or ΔVinc w[-]; P{hs::FLP}38/CyO; P{ovoD1-18}3L P{FRTw[hs]}2A (for genotypes with ΔVinc) males. Heat shocks were performed two times for 1 hr and 15 min each at 37°C at L1 and L2 larval stages. TalinIBS2-mCherry [31 (link)] was kindly provided by H.J. Bellen. The myosin heavy chain mutant used was Mhc[1] [45 (link)], kindly provided by S.I. Bernstein. IBS2-GFP recruitment to muscle attachment sites was performed with UAS::IBS2-GFP [15 (link)] expressed in muscles with P{Gal4-Mef2.R}3 (Bloomington Drosophila Stock Center).
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