![Role and regulation of HMGR-1 and the mevalonate pathway. (A) Left: Max intensity z-projected fluorescence and isosurface images showing the AC lysosomes (LMP-1::mNG) in a control and in a B0024.13 (dolichol synthesis) RNAi treated animal at the initiation of invasive protrusion formation. Right: Boxplot of AC lysosomal volume in control and B0024.13 RNAi treated animals (n = 20 control and 24 B0024.13 RNAi animals, ** P ≤ 0.01, Mann–Whitney U test). (B) Max intensity z-projected fluorescence and isosurface images showing the AC lysosomes (LMP-1::mNG) in a control and icmt-1 RNAi treated animal at the initiation of invasive protrusion formation. Boxplot of AC lysosome volume in control and icmt-1 RNAi treated animals (n = 23 control and 21 icmt-1 RNAi animals, ns [not statistically significant], P > 0.05, unpaired two-tailed Student’s t test). (C) Left: Sum intensity z-projection fluorescence images of GFP::CED-10 shows enrichment at the AC basal plasma membrane (arrow) in a control, an icmt-1, and a B0024.13 RNAi treated animal at the initiation of invasive protrusion formation. Right: Boxplot showing the basal level of GFP::CED-10 mean fluorescence intensity (n = 29 control, 22 B0024.13, and 21 icmt-1 RNAi animals, * P ≤ 0.05, Mann–Whitney U test). (D) Top: Single slice confocal images of mCherry::KDEL (magenta, ER marker), mNG::HMGR-1 (cyan), and overlay in the AC show overlap of mNG::HMGR-1 and mCherry::KDEL signals (white arrows) and a region of enriched mNG::HMGR-1 with no overlap (yellow arrows). Below: Insets highlighting region of increased mNG::HMGR-1 fluorescence where there is no mCherry::KDEL (yellow arrows; similar colocalization observed in = 8/8 animals). (E) Top: Single slice confocal images of mScarlet::PTS1 (magenta, peroxisome marker), mNG::HMGR-1 (cyan), and overlay in the AC shows overlap of enriched mNG::HMGR-1 regions and mScarlet::PTS1 (white arrows). Below: Insets highlighting region of overlap (arrows; similar colocalization observed in = 3/3 animals). (F) Left: Sum intensity z-projected fluorescence images of mNG::HMGR-1 in two ACs with mNG::HMGR-1 enriched at the invasive basal side (arrows). Right: mNG::HMGR-1 is not concentrated at the basal side of uterine cells (UCs, arrows, n = ≥10/10). (G) Left: Sum intensity z-projected fluorescence images of mNG::HMGR-1 in the AC (arrows show AC and enrichment at the invasive front) in a control and a sbp-1 RNAi treated animal. Right: Boxplot of mNG::HMGR-1 fluorescence intensity in the AC of control and sbp-1 RNAi animals (n = 26 control and 27 sbp-1 RNAi animals, unpaired two-tailed Student’s t test, ns [not statistically significant], P > 0.05, unpaired two-tailed Student’s t test). (H) Left: Sum intensity z-projected fluorescence images of mNG::HMGR-1 polarized (arrow) in a wild-type and mispolarized in an unc-6(ev400) mutant animal at the initiation of invasive protrusion formation. Bottom panels show spectral fluorescence-intensity maps, which display the minimum (L, low) and maximum (H, high) pixel value range of the acquired data. Right: Boxplot shows AC basal/apical ratio of mNG::HMGR-1 fluorescence intensity in wild-type and unc-6(ev400) animals (n = 48 control and 30 unc-6(ev400) animals, **** P ≤ 0.0001, Mann–Whitney U test). All data are from two or more replicates. Scale bars, 5 µm.](https://cdn.rupress.org/rup/content_public/journal/jcb/223/10/10.1083_jcb.202402035/2/jcb_202402035_figs4.png?Expires=1771484211&Signature=2boHDFFWDebNbN2lAFePQNg0NXenW5TX-bLg8MMIIHGly~D8COTRcJ4TA~Hz6g~zBr2e84OBG2er91sMrkW5Q2uyH4ZmbSmJTNRDpHDl5lvSX7NDzSEb46aOCNInF3C0U0cppqQDABeR2u-hgFqzip8t9VolsqfggCp4R3jtRnUk0mEgzfo2j3tQFIXmsbgIFJQ62wo0YmM2DYiTzTROLoRJE0pxNTcOKvqfrXf3HekfSLzHpvwdy17e5YcqlL~kowOvWmY4YycrkbesLt1Ndu8RkzCDUg-V2j~P-zUwA0OPocSrOqoyNZGxSq~iYrfz4-J7JvPHcnDwDGbSr1jmrg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Role and regulation of HMGR-1 and the mevalonate pathway. (A) Left: Max intensity z-projected fluorescence and isosurface images showing the AC lysosomes (LMP-1::mNG) in a control and in a B0024.13 (dolichol synthesis) RNAi treated animal at the initiation of invasive protrusion formation. Right: Boxplot of AC lysosomal volume in control and B0024.13 RNAi treated animals (n = 20 control and 24 B0024.13 RNAi animals, ** P ≤ 0.01, Mann–Whitney U test). (B) Max intensity z-projected fluorescence and isosurface images showing the AC lysosomes (LMP-1::mNG) in a control and icmt-1 RNAi treated animal at the initiation of invasive protrusion formation. Boxplot of AC lysosome volume in control and icmt-1 RNAi treated animals (n = 23 control and 21 icmt-1 RNAi animals, ns [not statistically significant], P > 0.05, unpaired two-tailed Student’s t test). (C) Left: Sum intensity z-projection fluorescence images of GFP::CED-10 shows enrichment at the AC basal plasma membrane (arrow) in a control, an icmt-1, and a B0024.13 RNAi treated animal at the initiation of invasive protrusion formation. Right: Boxplot showing the basal level of GFP::CED-10 mean fluorescence intensity (n = 29 control, 22 B0024.13, and 21 icmt-1 RNAi animals, * P ≤ 0.05, Mann–Whitney U test). (D) Top: Single slice confocal images of mCherry::KDEL (magenta, ER marker), mNG::HMGR-1 (cyan), and overlay in the AC show overlap of mNG::HMGR-1 and mCherry::KDEL signals (white arrows) and a region of enriched mNG::HMGR-1 with no overlap (yellow arrows). Below: Insets highlighting region of increased mNG::HMGR-1 fluorescence where there is no mCherry::KDEL (yellow arrows; similar colocalization observed in = 8/8 animals). (E) Top: Single slice confocal images of mScarlet::PTS1 (magenta, peroxisome marker), mNG::HMGR-1 (cyan), and overlay in the AC shows overlap of enriched mNG::HMGR-1 regions and mScarlet::PTS1 (white arrows). Below: Insets highlighting region of overlap (arrows; similar colocalization observed in = 3/3 animals). (F) Left: Sum intensity z-projected fluorescence images of mNG::HMGR-1 in two ACs with mNG::HMGR-1 enriched at the invasive basal side (arrows). Right: mNG::HMGR-1 is not concentrated at the basal side of uterine cells (UCs, arrows, n = ≥10/10). (G) Left: Sum intensity z-projected fluorescence images of mNG::HMGR-1 in the AC (arrows show AC and enrichment at the invasive front) in a control and a sbp-1 RNAi treated animal. Right: Boxplot of mNG::HMGR-1 fluorescence intensity in the AC of control and sbp-1 RNAi animals (n = 26 control and 27 sbp-1 RNAi animals, unpaired two-tailed Student’s t test, ns [not statistically significant], P > 0.05, unpaired two-tailed Student’s t test). (H) Left: Sum intensity z-projected fluorescence images of mNG::HMGR-1 polarized (arrow) in a wild-type and mispolarized in an unc-6(ev400) mutant animal at the initiation of invasive protrusion formation. Bottom panels show spectral fluorescence-intensity maps, which display the minimum (L, low) and maximum (H, high) pixel value range of the acquired data. Right: Boxplot shows AC basal/apical ratio of mNG::HMGR-1 fluorescence intensity in wild-type and unc-6(ev400) animals (n = 48 control and 30 unc-6(ev400) animals, **** P ≤ 0.0001, Mann–Whitney U test). All data are from two or more replicates. Scale bars, 5 µm.
