Figure S5. NPR-12 acts cell autonomously to regulate dendritedegeneration, Related to Figure 4 and Figure 6(A) Quantification of PVD dendrite degeneration in the mutants of candidatereceptors for NLP-29:
npr-2,
npr-32,
pdfr-1, and
dmsr-1. One-way ANOVA,followed by Fisher's LSD post-hoc test, was used to detect the differencesbetween each mutant, control and
nlp-29, for each day. Mutant versus control:#(black) p < 0.05, ##(black) p < 0.001. Mutant versus
nlp-29: #(pink) p < 0.05,##(pink) p < 0.001. Non-significant comparisons are not indicated in the figure.(B) Quantification of PVD dendrite degeneration in animals with PVD-specificRNAi knock-down of
lat-1, C17H11.1 and T21H3.5. Empty vector was used asnegative control. Student's t-test showed no difference between control animalsand
lat-1, C17H11.1 or T21H3.5 at any time point.(C) A Pnpr-12::NPR-12::GFP translational reporter strain was used to visualizeNPR-12 protein expression in PVD neurons. A 2.2 kb fragment upstream of thestart codon of
npr-12 was used as the
npr-12 promoter. White arrow: PVD cellbody; red arrow: PVD dendrite; yellow arrow: initial segment of PVD axon. Scalebar: 10 um.(D) A Pnpr-12::GFP strain was used to visualize the gene expression pattern ofnpr-12. A 2.2 kb fragment upstream of the start codon of
npr-12 was used as thenpr-12 promoter. Representative images of mid-section, head region, and tailregion of an animal are shown in the top, middle and bottom panels, respectively.The white arrow in the top image points at the PVD cell body. The middle imageshows
npr-12 expression in FLP neurons (white arrow points at FLP cell body), and the bottom image demonstrates
npr-12 expression in PVC neurons andpossibly in PHA neurons as well. The neurons in yellow color are AIY neuronsfrom expression of co-injection marker Pttx-3::RFP. Scale bars: 50 um.(E) RT-qPCR quantification of
npr-12 mRNA levels in control animals. One-wayANOVA analysis showed no significant differences over the course of aging.(F) Quantification of PVD dendrite degeneration in animals with epidermisspecific or PVD-specific RNAi knockdown of
npr-12. Empty vectors were used asnegative control.
npr-12 (PVD) RNAi versus Control (PVD): #p < 0.05, ##p <0.001. Student's t-test.(G) Representative images of aging-associated dendrite degeneration of amphidsensory neurons, in control animals (Parl-13::GFP) as well as animals withectopic expression of
npr-12 in amphid sensory neurons driven by the
arl-13promoter (Parl-13::
npr-12). White arrows point at beading, and red arrows pointat vacuoles, both considered dendrite degeneration phenotypes. Scale bar: 50um.(H) Quantification of dendrite degeneration of amphid sensory neurons, in control(Parl-13::GFP) and Parl-13::
npr-12 animals, on Day 1 and Day 9. One-wayANOVA, followed by Fisher's LSD post-hoc test. ## p < 0.001.Data are represented as mean +- SEM. Non-significant comparisons are notindicated in the figure.