Figure 1. Wounding triggers PtdIns4 P... | Rockefeller University Press

Figure 1.

$Wounding triggers PtdIns4P accumulation and PtdIns(4,5)P2generation at the wound site. (A) Illustration showing C. elegans epidermal cell hyp7 as a model for membrane repair. In this study, membrane components PtdIns(4,5)P2 were labeled by the PH domain from PLC-δ1, and PtdIns4P were labeled by the P4M domain from L. pneumophila SidM. (B) Time-lapse images showing the accumulation of PH::GFP at the wound site. “W” indicates the wound site. Pcol-19-PH::GFP (zjuSi175) transgenic animals were used for wounding and imaging. Scale bar: 10 µm. (C) Representative HiS-SiM single-plane images of PH::GFP before and after wounding. Pcol-19-PH::GFP(zjuSi175) transgenic animals were used for wounding and imaging. Scale bar: 10 µm. (D) Representative confocal time-lapse images of the recruitment of mKate2::P4M and PH::GFP between 5 to 90 min after wounding. Pcol-19-PH::GFP(zjuSi175);Pcol-19-mKate2::P4M (zjuSi333) transgenic animals were used for wounding and imaging. Scale bar: 10 µm. (E) Line graph showing the intensity change (ΔFw/Fuw) of mKate2::P4M and PH::GFP at different time points after wounding. The plot indicates the mean ± SD (n = 6 for mKate2::P4M curve and 8 for PH::GFP curve) at each time point. (F) Fluorescent recovery after photobleaching (FRAP) analysis of the recovery capability of mKate2::P4M and PH::GFP before and 1 h after wounding. Pcol-19-PH::GFP(zjuSi175);Pcol-19::mKate2::P4M(zjuSi333) transgenic animals were used for wounding and imaging. Scale bar: 10 µm. (G) Mobile fraction (Fm) analysis of FRAP of mKate2::P4M and PH::GFP in F. Error bars represent the mean value ± SD (n = 27, 18, 27, 18 from the left to the right), Mann–Whitney test, ***P < 0.001. See also Fig. S1; and Videos 1, 2, 3, 4, 5, 6, and 7.$

Wounding triggers PtdIns4P accumulation and PtdIns(4,5)P ₂ generation at the wound site. (A) Illustration showing C. elegans epidermal cell hyp7 as a model for membrane repair. In this study, membrane components PtdIns(4,5)P2 were labeled by the PH domain from PLC-δ1, and PtdIns4P were labeled by the P4M domain from L. pneumophila SidM. (B) Time-lapse images showing the accumulation of PH::GFP at the wound site. “W” indicates the wound site. Pcol-19-PH::GFP (zjuSi175) transgenic animals were used for wounding and imaging. Scale bar: 10 µm. (C) Representative HiS-SiM single-plane images of PH::GFP before and after wounding. Pcol-19-PH::GFP(zjuSi175) transgenic animals were used for wounding and imaging. Scale bar: 10 µm. (D) Representative confocal time-lapse images of the recruitment of mKate2::P4M and PH::GFP between 5 to 90 min after wounding. Pcol-19-PH::GFP(zjuSi175);Pcol-19-mKate2::P4M (zjuSi333) transgenic animals were used for wounding and imaging. Scale bar: 10 µm. (E) Line graph showing the intensity change (ΔF_w/F_uw) of mKate2::P4M and PH::GFP at different time points after wounding. The plot indicates the mean ± SD (n = 6 for mKate2::P4M curve and 8 for PH::GFP curve) at each time point. (F) Fluorescent recovery after photobleaching (FRAP) analysis of the recovery capability of mKate2::P4M and PH::GFP before and 1 h after wounding. Pcol-19-PH::GFP(zjuSi175);Pcol-19::mKate2::P4M(zjuSi333) transgenic animals were used for wounding and imaging. Scale bar: 10 µm. (G) Mobile fraction (F_m) analysis of FRAP of mKate2::P4M and PH::GFP in F. Error bars represent the mean value ± SD (n = 27, 18, 27, 18 from the left to the right), Mann–Whitney test, ***P < 0.001. See also Fig. S1; and Videos 1, 2, 3, 4, 5, 6, and 7.