![POD-2, FASN-1, ELO-1, LPIN-1, and SMS-1 AC expression from P6.p 1-cell to 4-cell stage. (A) Left: Sum intensity z-projected fluorescence images of endogenously tagged GFP::POD-2, FASN-1::mNG, ELO-1::mNG, mNG::LPIN-1, and SMS-1::mNG in the AC (arrows and white dotted lines) from the P6.p 1-cell to 4-cell stages. Right: Boxplots show the mean fluorescence intensity of each protein in the AC (comparisons of P6.p 1-cell and 2–4-cell stages, [POD-2] n = 11 1-cell, 10 2-cell, 10 2–4-cell, and 11 4-cell stage animals; [FASN-1] n = 11 1-cell, 12 2-cell, 10 2–4-cell, and 10 4-cell stage animals; [ELO-1] n = 14 1-cell, 13 2-cell, 9 2–4-cell, and 13 4-cell stage animals; [LPIN-1] n = 8 1-cell, 15 2-cell, 10 2–4-cell, and 14 4-cell stage animals; [SMS-1] n = 10 1-cell, 10 2-cell, 5 2–4-cell, and 10 4-cell stage animals, * P ≤ 0.05, *** P ≤ 0.001, ns [not statistically significant], P > 0.05, Brown-Forsythe and Welch ANOVA tests followed by Dunnett’s T3 multiple comparisons test and Kruskal–Wallis test followed by Dunn’s multiple comparisons test; comparisons of uterine cell [UC] to AC at P6.p 4-cell stage, [POD-2] n = 11 animals; [FASN-1] n = 10 animals; [ELO-1] n = 13 animals; [LPIN-1] n = 14 animals; [SMS-1] n = 10 animals, * P ≤ 0.05, ** P ≤ 0.001, *** P ≤ 0.0001, P = 0.10, Student’s t test). (B) Top: Single slice confocal images of mCherry::KDEL (magenta), ELO-1::mNG (cyan) and overlay in the AC show tight overlap (arrows). Bottom: Insets highlighting the overlap in the basal region (arrows; similar colocalization observed in n = 10/10 animals). (C) Top: Max intensity z-projected fluorescence images of AMAN-2::mScarlet (magenta, Golgi), SMS-1::mNG (cyan), and overlay in the AC reveal colocalization (arrows). The average percentage ± SD of colocalization of SMS-1::mNG puncta with AMAN-2::mScarlet with is indicated (n = 6 animals). Below: Insets highlight AMAN-2::mScarlet (magenta) and SMS-1::mNG (cyan) colocalization (arrows). (D) Left: Sum intensity z-projections of ZMP-1::mNG fluorescence showing the invasive membrane enrichment (arrow) in a control and a ppk-3 RNAi treated animal at the initiation of invasive protrusion formation. Right: Boxplot showing the basal ZMP-1::mNG mean fluorescence signal in control and ppk-3 RNAi treated animals (n = 8 control and 9 ppk-3 RNAi animals, P ≤ 0.05, 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.202402035/1/jcb_202402035_figs3.png?Expires=1768355738&Signature=DoKHgNDkAyt8xPK76bYshuBdnRbi-PHX4ZHBCKj8knamUaOdMBIEp6Y3x4jjpDDjA9EV--tBBbCEeaMz5TpFS66h~U14vnyn-uzQ7XD-FwZl1~egFtYbYjtRaFyPJj79k51LpPDQCuiRIS3KgE0yydU6d9Nj4NIFiF8GwciDpuBab0mcb59lQ-NAx5C0Rn7weHty2I2qDYtvHa5ZqAn4JTJmivlrBC-7m0eF0hRarPCqlvQTtXhYplurQiD87zms6eBGMdi2vvUvBft9-xOJNIqxJ~P-xXX~HhV8CiXmHOxEW9-FPS~JXYjtJ757n6GcW6l-BeWkHRsO2B5a0w2XKQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
POD-2, FASN-1, ELO-1, LPIN-1, and SMS-1 AC expression from P6.p 1-cell to 4-cell stage. (A) Left: Sum intensity z-projected fluorescence images of endogenously tagged GFP::POD-2, FASN-1::mNG, ELO-1::mNG, mNG::LPIN-1, and SMS-1::mNG in the AC (arrows and white dotted lines) from the P6.p 1-cell to 4-cell stages. Right: Boxplots show the mean fluorescence intensity of each protein in the AC (comparisons of P6.p 1-cell and 2–4-cell stages, [POD-2] n = 11 1-cell, 10 2-cell, 10 2–4-cell, and 11 4-cell stage animals; [FASN-1] n = 11 1-cell, 12 2-cell, 10 2–4-cell, and 10 4-cell stage animals; [ELO-1] n = 14 1-cell, 13 2-cell, 9 2–4-cell, and 13 4-cell stage animals; [LPIN-1] n = 8 1-cell, 15 2-cell, 10 2–4-cell, and 14 4-cell stage animals; [SMS-1] n = 10 1-cell, 10 2-cell, 5 2–4-cell, and 10 4-cell stage animals, * P ≤ 0.05, *** P ≤ 0.001, ns [not statistically significant], P > 0.05, Brown-Forsythe and Welch ANOVA tests followed by Dunnett’s T3 multiple comparisons test and Kruskal–Wallis test followed by Dunn’s multiple comparisons test; comparisons of uterine cell [UC] to AC at P6.p 4-cell stage, [POD-2] n = 11 animals; [FASN-1] n = 10 animals; [ELO-1] n = 13 animals; [LPIN-1] n = 14 animals; [SMS-1] n = 10 animals, * P ≤ 0.05, ** P ≤ 0.001, *** P ≤ 0.0001, P = 0.10, Student’s t test). (B) Top: Single slice confocal images of mCherry::KDEL (magenta), ELO-1::mNG (cyan) and overlay in the AC show tight overlap (arrows). Bottom: Insets highlighting the overlap in the basal region (arrows; similar colocalization observed in n = 10/10 animals). (C) Top: Max intensity z-projected fluorescence images of AMAN-2::mScarlet (magenta, Golgi), SMS-1::mNG (cyan), and overlay in the AC reveal colocalization (arrows). The average percentage ± SD of colocalization of SMS-1::mNG puncta with AMAN-2::mScarlet with is indicated (n = 6 animals). Below: Insets highlight AMAN-2::mScarlet (magenta) and SMS-1::mNG (cyan) colocalization (arrows). (D) Left: Sum intensity z-projections of ZMP-1::mNG fluorescence showing the invasive membrane enrichment (arrow) in a control and a ppk-3 RNAi treated animal at the initiation of invasive protrusion formation. Right: Boxplot showing the basal ZMP-1::mNG mean fluorescence signal in control and ppk-3 RNAi treated animals (n = 8 control and 9 ppk-3 RNAi animals, P ≤ 0.05, Mann–Whitney U test). All data are from two or more replicates. Scale bars, 5 µm.