PVF-1/VER signaling acts with the VPS-35/retromer recycling pathway to regulate GLR-1 surface levels.
(A-E) Representative images and quantification of the SEP and mCherry fluorescence and the SEP/mCherry ratio of the indicated genotypes are shown. Images were acquired at low laser intensity to avoid saturation. Note Y-axis log scale. Mean ± SEM are shown (n ≥ 20 worms from 3 experiments). *p ≤ 0.05 ANOVA followed by Tukey’s multiple comparison test. (F) Quantification of the SEP and mCherry fluorescence or the SEP/mCherry ratio of the indicated genotypes are shown. Mean ± SEM are shown (n ≥ 30 worms from ≥ 3 experiments). *p ≤ 0.05, ANOVA, Tukey’s multiple comparison test. Values that differ significantly from wild type are indicated by asterisks above each bar, whereas other comparisons are marked by brackets. (G) Representative Dual FRAP images showing pre-bleach, bleach, and 10 minute recovery images of SEP::mCherry::GLR-1 (akIs201) in a region (outlined by a dotted line) in the VNC of L4 stage larval wild type worms. (H) Quantification of Dual FRAP (bleach at 0 min) of SEP∷mCherry∷GLR-1 in the VNC of WT, ver-1, vps-35 or lin-10 worms. A 50% smaller ROI that was centered within the larger bleach ROI shown was used for quantification to minimize contributions from laterally diffusing receptors. At 10 min, lin-10 SEP vs WT SEP, *p < 0.001. Two-way repeated measures ANOVA, followed by Tukey’s multiple comparison test. Merge images have been false colored magenta (mCherry) and green (SEP).