Picture from Hoege C et al. (2010) Curr Biol "LGL can partition the cortex of one-cell Caenorhabditis elegans embryos into two ...."
Figure 1. LGL Interacts with Anterior PAR Complex Proteins but Localizes to the Posterior Cortex. (A) LGL and PKC-3 interact with Caenorhabditis elegans PAR-6. Anti-PAR-6 immunoprecipitations were made from wild-type C. elegans worms. Proteins were stained with Coomassie, and specific bands were excised and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry [15]. PAR-6 (36 kDa) stains only weakly by Coomassie. The 80 kDa band contained PKC-3, the 120 kDa band contained the protein of F56F10.4/lgl, and a third band at 180 kDa contained the protein of W07E11.1. * denotes IgG bands.(B) GFP-LGL interacts with PKC-3 and PAR-6 in C. elegans embryos. Immunoblots were made from anti-GFP immunoprecipitation from GFP-LGL-expressing embryos.(C) The LGL complex is sensitive to phosphatase treatment. Anti-GFP immunoprecipitations from GFP-LGL-expressing embryos were left untreated (-) or treated (+) with calf intestine phosphatase (CIP). Proteins were detected by immunoblot.(D) GFP-LGL localizes to the posterior cortex after polarity onset. Images are from time-lapse recording. Times are relative to nuclear envelope breakdown (0 s). Top row: differential interference contrast images; bottom row: GFP-LGL fluorescence. Note the blow-up of the anterior and posterior cortex region.(E) LGL colocalizes with PAR-2. mcherry-PAR-2 and GFP-LGL from a double fluorescent cell line colocalize on the posterior cortex of the one-cell embryo.(F) Immunofluorescence of LGL in embryos treated with control(RNAi), lgl(RNAi), or GFP-LGL-expressing embryos, or GFP-LGL embryos silenced only for the transgene gfp::lgl by gfp(RNAi) (DNA, blue; microtubules, green; LGL, red). All images were taken with identical exposure conditions and processed identically. For quantifications, see Figure S1F.