Fig. 2: Aberrant recruitment of proteins involved in actomyosin contraction to PI(4,5)P2-positive endosomal membranes in the absence of SMP proteins. | Nature Communications

Fig. 2: Aberrant recruitment of proteins involved in actomyosin contraction to PI(4,5)P2-positive endosomal membranes in the absence of SMP proteins.

From: PDZD-8 and TEX-2 regulate endosomal PI(4,5)P2 homeostasis via lipid transport to promote embryogenesis in C. elegans

Fig. 2: Aberrant recruitment of proteins involved in actomyosin contraction to PI(4,5)P2-positive endosomal membranes in the absence of SMP proteins.The alternative text for this image may have been generated using AI.

a Left: A representative live spinning disc confocal (SDC) image of an equatorial plane of an early embryo from a mutant lacking all the four SMP proteins [quadruple knock-out: QKO (pdzd-8; tex-2; tmem-24; esyt-2)], co-expressing PI(4,5)P2 biosensor (mCherry::PHPLCδ1) and NMY-2-tagged with GFP (NMY-2::GFP). The image is from the polarity establishment phase. Right: Magnified insets, showing PI(4,5)P2-positive puncta, as determined by the presence of mCherry::PHPLCδ1, that are outlined by indicated white dotted boxes. Scale bar, 10 μm. b Time-lapse SDC images of a representative PI(4,5)P2-positive puncta from an early embryo from QKO, co-expressing mCherry::PHPLCδ1 and NMY-2::GFP, over the period of 2 min. White arrowheads indicate the positions of the PI(4,5)P2-positive puncta. Note the extensive overlap of NMY-2::GFP signals with the PI(4,5)P2-positive puncta. Scale bars, 5 μm. c Time-lapse images of representative NMY-2::GFP puncta from an early embryo from QKO over the period of 50 s. Top panel: images from SDC microscopy. Bottom panel: images from SDC structure illumination microscopy (SDC-SIM). Note the lumen inside the NMY-2::GFP puncta in SDC-SIM images. Scale bars, 5 μm. d, e Left: Live SDC-SIM images of early embryos from QKO, expressing NMY-2::GFP together with either d wrmScarlet-tagged ANI-1 (wrmScarlet::ANI-1) or e lifeACT-tagged with RFP (lifeACT::RFP), showing representative NMY-2::GFP puncta that are co-localized with the marker proteins as indicated. Right: Line scan profiles of NMY-2::GFP (green) and either d wrmScarlet::ANI-1 or e lifeACT::RFP (red) fluorescence signals along the white dotted lines as indicated. Scale bars, 1 μm. f, g Left: Live SDC-SIM images of early embryos from QKO, expressing wrmScarlet::ANI-1 together with either f GFP-tagged RAB-7 (GFP::RAB-7) or g GFP-tagged RAB-11.1 (GFP::RAB-11.1), showing representative wrmScarlet::ANI-1 puncta that are co-localized with the marker proteins as indicated. Right: Line scan profiles of wrmScarlet::ANI-1 (red) and either f GFP::RAB-7 or g GFP::RAB-11.1 (green) fluorescence signals along the white dotted lines as indicated. Scale bars, 1 μm.

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