Figure 5.
SMS-1 promotes formation of lysosome stores and localization of UNC-40 and ZMP-1. (A) Left: Maximum intensity z-projected fluorescence images showing the AC (cyan, mCherry::PLCδPH) in a control and an AC-specific sms-1 RNAi treated animal (BM, orange dashed lines) with a small BM breach observed corresponding to the reduced protrusion after loss of sms-1 (middle, bracket, observed in n = 5/5 sms-1 RNAi animals). Isosurfaces of the AC invasive protrusion (below, arrowheads) at the time of maximum protrusion volume (see Video 6). Right: Boxplot of the maximum invasive protrusion volume in control and sms-1 RNAi treated animals (n = 5 control and 5 sms-1 RNAi animals, ** P ≤ 0.01, Mann–Whitney U test). (B) Left: Max intensity z-projected fluorescence images and isosurfaces showing the AC lysosomes (LMP-1::mNG) in control and sms-1 RNAi-treated animal at the initiation of protrusion formation. Right: Boxplot of the AC lysosome volume in control and sms-1 RNAi treated animals (n = 24 control and 23 sms-1 RNAi animals, ** P ≤ 0.01, unpaired two-tailed Student’s t test). (C) Left: Sum intensity z-projected fluorescence images of GFP::CED-10 and GFP::MIG-2. Arrows show enrichment at the AC invasive membrane in a control and sms-1 RNAi treated animals at the time of invasive protrusion initiation. Right: Boxplots show the basal/apical ratio of GFP::CED-10 and GFP::MIG-2 fluorescence intensity in control and sms-1 RNAi treated animals ([CED-10] n = 16 control and 15 sms-1 RNAi animals; [MIG-2] n = 12 control and 11 sms-1 RNAi animals, ns [not statistically significant], P > 0.05, Mann–Whitney U test). (D) Left: Sum intensity fluorescence images of the GPI-anchored matrix metalloproteinase ZMP-1::mNG showing the AC invasive membrane (arrows) in a control and a sms-1 RNAi treated animal at the initiation of invasive protrusion formation. Right: Boxplot showing the AC basal/apical ratio of ZMP-1::mNG fluorescence intensity in control and zmp-1 RNAi treated animals (n = 16 control and 13 sms-1 RNAi animals, * P ≤ 0.05, unpaired two-tailed Student’s t test). (E) Left: Ventral view of the AC-BM interface (BM visualized with LAM-1::mCherry) showing UNC-40::GFP enrichment (arrows) at the initial BM breach (orange dotted lines) in a control and a sms-1 treated RNAi animal. Right: Boxplot shows UNC-40::GFP mean fluorescence intensity at the initial BM breach (n = 6 control and 6 sms-1 RNAi animals, * P ≤ 0.05, Mann–Whitney U test). (F) A schematic diagram summarizing the roles of the transcription factor SBP-1, the fatty acid synthesis enzymes POD-2 and FASN-1, and phospholipid synthesizing LPIN-1 and sphingomyelin catalyzing SMS-1 in AC invasive protrusion formation. All data in the figure are from two or more replicates. Scale bars, 5 µm.

SMS-1 promotes formation of lysosome stores and localization of UNC-40 and ZMP-1. (A) Left: Maximum intensity z-projected fluorescence images showing the AC (cyan, mCherry::PLCδPH) in a control and an AC-specific sms-1 RNAi treated animal (BM, orange dashed lines) with a small BM breach observed corresponding to the reduced protrusion after loss of sms-1 (middle, bracket, observed in n = 5/5 sms-1 RNAi animals). Isosurfaces of the AC invasive protrusion (below, arrowheads) at the time of maximum protrusion volume (see Video 6). Right: Boxplot of the maximum invasive protrusion volume in control and sms-1 RNAi treated animals (n = 5 control and 5 sms-1 RNAi animals, ** P ≤ 0.01, Mann–Whitney U test). (B) Left: Max intensity z-projected fluorescence images and isosurfaces showing the AC lysosomes (LMP-1::mNG) in control and sms-1 RNAi-treated animal at the initiation of protrusion formation. Right: Boxplot of the AC lysosome volume in control and sms-1 RNAi treated animals (n = 24 control and 23 sms-1 RNAi animals, ** P ≤ 0.01, unpaired two-tailed Student’s t test). (C) Left: Sum intensity z-projected fluorescence images of GFP::CED-10 and GFP::MIG-2. Arrows show enrichment at the AC invasive membrane in a control and sms-1 RNAi treated animals at the time of invasive protrusion initiation. Right: Boxplots show the basal/apical ratio of GFP::CED-10 and GFP::MIG-2 fluorescence intensity in control and sms-1 RNAi treated animals ([CED-10] n = 16 control and 15 sms-1 RNAi animals; [MIG-2] n = 12 control and 11 sms-1 RNAi animals, ns [not statistically significant], P > 0.05, Mann–Whitney U test). (D) Left: Sum intensity fluorescence images of the GPI-anchored matrix metalloproteinase ZMP-1::mNG showing the AC invasive membrane (arrows) in a control and a sms-1 RNAi treated animal at the initiation of invasive protrusion formation. Right: Boxplot showing the AC basal/apical ratio of ZMP-1::mNG fluorescence intensity in control and zmp-1 RNAi treated animals (n = 16 control and 13 sms-1 RNAi animals, * P ≤ 0.05, unpaired two-tailed Student’s t test). (E) Left: Ventral view of the AC-BM interface (BM visualized with LAM-1::mCherry) showing UNC-40::GFP enrichment (arrows) at the initial BM breach (orange dotted lines) in a control and a sms-1 treated RNAi animal. Right: Boxplot shows UNC-40::GFP mean fluorescence intensity at the initial BM breach (n = 6 control and 6 sms-1 RNAi animals, * P ≤ 0.05, Mann–Whitney U test). (F) A schematic diagram summarizing the roles of the transcription factor SBP-1, the fatty acid synthesis enzymes POD-2 and FASN-1, and phospholipid synthesizing LPIN-1 and sphingomyelin catalyzing SMS-1 in AC invasive protrusion formation. All data in the figure are from two or more replicates. Scale bars, 5 µm.

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