Figure 6.
HMGR-1 promotes lysosome stores, GTPase accumulation, and invasive protrusion formation. (A) A schematic diagram showing the mevalonate pathway in the AC (see Fig. S1 B for pathway details, Table S2 for RNAi screen). (B) Left: Maximum intensity z-projected fluorescence images showing the AC (cyan, mCherry::PLCδPH) protrusion (arrowheads) in a control and an AC-specific hmgr-1 RNAi treated animal (BM, orange dashed lines) with a small BM breach observed corresponding to the reduced protrusion after loss of hmgr-1 (middle, bracket, observed in n = 7/7 hmgr-1 RNAi animals). Isosurfaces of the AC invasive protrusion (below, arrowheads) at the time of maximum protrusion volume (see Video 7). Right: Boxplot of the maximum invasive protrusion volume in control and hmgr-1 RNAi treated animals (n = 9 control and 7 hmgr-1 RNAi animals, ** P ≤ 0.01, Mann–Whitney U test). (C) Left: Max intensity z-projected fluorescence images and isosurfaces showing the AC lysosomes (LMP-1::mNG) in a control and an hmgr-1 RNAi-treated animal at the initiation of protrusion formation. Right: Boxplot of the AC lysosome volume of control and hmgr-1 RNAi animals (n = 17 control and 18 hmgr-1 RNAi animals, *** P ≤ 0.001, unpaired two-tailed Student’s t test). (D) Top: Sum intensity z-projected images showing invasive polarization (arrows) of GFP::CED-10, GFP::MIG-2, GFP::CAAX, and mNG::RAP-1 at the AC plasma membrane in control and hmgr-1 RNAi animals at the initiation of invasive protrusion formation. Bottom: Boxplots showing the AC basal/apical ratio of each protein’s fluorescence intensity in control and hmgr-1 RNAi-treated animals ([CED-10] n = 23 control and 23 hmgr-1 RNAi animals; [MIG-2] n = 13 control and 13 hmgr-1 RNAi animals; [CAAX] n = 16 control and 16 hmgr-1 RNAi animals; [RAP-1] n = 17 control and 18 hmgr-1 RNAi animals, ** P ≤ 0.01, **** P ≤ 0.0001, unpaired two-tailed Student’s t test). (E) Top: Sum intensity projected images of ZMP-1::mNG shows ZMP-1 enrichment at the AC basal invasive plasma membrane (arrows) in a control and an hmgr-1 RNAi-treated animal at the initiation of invasive protrusion formation. Bottom: Boxplot showing the AC basal/apical ratio of ZMP-1::mNG fluorescence intensity in control and hmgr-1 RNAi treated animals (n = 14 control and 16 hmgr-1 RNAi animals, ns [not statistically significant], P > 0.05, unpaired two-tailed Student’s t test). All data are from two or more replicates. Scale bars, 5 µm.

HMGR-1 promotes lysosome stores, GTPase accumulation, and invasive protrusion formation. (A) A schematic diagram showing the mevalonate pathway in the AC (see Fig. S1 B for pathway details, Table S2 for RNAi screen). (B) Left: Maximum intensity z-projected fluorescence images showing the AC (cyan, mCherry::PLCδPH) protrusion (arrowheads) in a control and an AC-specific hmgr-1 RNAi treated animal (BM, orange dashed lines) with a small BM breach observed corresponding to the reduced protrusion after loss of hmgr-1 (middle, bracket, observed in n = 7/7 hmgr-1 RNAi animals). Isosurfaces of the AC invasive protrusion (below, arrowheads) at the time of maximum protrusion volume (see Video 7). Right: Boxplot of the maximum invasive protrusion volume in control and hmgr-1 RNAi treated animals (n = 9 control and 7 hmgr-1 RNAi animals, ** P ≤ 0.01, Mann–Whitney U test). (C) Left: Max intensity z-projected fluorescence images and isosurfaces showing the AC lysosomes (LMP-1::mNG) in a control and an hmgr-1 RNAi-treated animal at the initiation of protrusion formation. Right: Boxplot of the AC lysosome volume of control and hmgr-1 RNAi animals (n = 17 control and 18 hmgr-1 RNAi animals, *** P ≤ 0.001, unpaired two-tailed Student’s t test). (D) Top: Sum intensity z-projected images showing invasive polarization (arrows) of GFP::CED-10, GFP::MIG-2, GFP::CAAX, and mNG::RAP-1 at the AC plasma membrane in control and hmgr-1 RNAi animals at the initiation of invasive protrusion formation. Bottom: Boxplots showing the AC basal/apical ratio of each protein’s fluorescence intensity in control and hmgr-1 RNAi-treated animals ([CED-10] n = 23 control and 23 hmgr-1 RNAi animals; [MIG-2] n = 13 control and 13 hmgr-1 RNAi animals; [CAAX] n = 16 control and 16 hmgr-1 RNAi animals; [RAP-1] n = 17 control and 18 hmgr-1 RNAi animals, ** P ≤ 0.01, **** P ≤ 0.0001, unpaired two-tailed Student’s t test). (E) Top: Sum intensity projected images of ZMP-1::mNG shows ZMP-1 enrichment at the AC basal invasive plasma membrane (arrows) in a control and an hmgr-1 RNAi-treated animal at the initiation of invasive protrusion formation. Bottom: Boxplot showing the AC basal/apical ratio of ZMP-1::mNG fluorescence intensity in control and hmgr-1 RNAi treated animals (n = 14 control and 16 hmgr-1 RNAi animals, ns [not statistically significant], P > 0.05, unpaired two-tailed Student’s t test). All data are from two or more replicates. Scale bars, 5 µm.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal