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. 2023 Aug 31;21(8):e3002276. doi: 10.1371/journal.pbio.3002276

Fig 7. Epidermal UNC-31 is involved in the epidermal ribosome biogenesis inhibition-mediated growth quiescence.

Fig 7

(A) Synchronized embryos of the strain with a ribosomal protein gene null, rps-23(0), and M cell lineage marker (hlh-8p::GFP) were grown on the plates seeded with RNAi bacteria targeting the unc-31 gene or control for 3 days. Larvae with divided M cells were assessed. Reducing unc-31 expression by RNAi increased the percentage of larvae with divided M cells. Data were obtained from 9 independent experiments with at least 15 animals for each. Statistical significance was determined using an independent t test. (B) Embryos expressing degron::GFP-integrated RPOA-2 and TIR1 in the epidermis were treated with and without 1 mM IAA and fed by RNAi bacteria targeting unc-31 gene or control for 3 days. Without IAA treatment, RNAi unc-31 did not affect worm body length (left). With IAA treatment, animals fed by unc-31 RNAi bacteria grew larger compared to control. (C) A null allele of unc-31 mutant, unc-31(e928), was crossed to an inducible epidermal ribosome biogenesis strain (grwd-1::degron::GFP; col-10p::TIR1). unc-31(e928) mutants grew smaller compared to wild type, when epidermal GRWD-1 was depleted (+IAA). (D-F) Tissue-specific RNAi strains were crossed with an inducible epidermal ribosome biogenesis inhibition strain to detect the function of UNC-31 in different tissues. (D) Neuron-specific unc-31 RNAi animals (grwd-1::degron::GFP; col-10p::TIR1; sid-1(pk3321); unc-119p::sid-1) grew smaller without epidermal new ribosomes (+IAA). (E) Reducing unc-31 expression in muscle (grwd-1::degron::GFP; col-10p::TIR1; rde-1(ne300); myo-3p::rde-1) reduced body length with epidermal ribosome biogenesis inhibition (+IAA). (F) Reducing epidermal unc-31 expression (degron::GFP:: rpoa-2; col-10p::TIR1; rde-1(ne219); wrt-2p::rde-1, -IAA) did not change worm body length (left). When the epidermal ribosome biogenesis was inhibited (+IAA), animals with reduced unc-31 expression in epidermis resulted in a larger body length. Synchronized embryos were incubated for 3 days. Animals were immobilized using 0.5% 1-phenoxy-2-propanol. Each 5× image was analyzed by a custom MATLAB script (S1 Text). Data with IAA treatment were obtained from 3 independent experiments with at least 18 animals for each; data without IAA treatment were analyzed from 26 animals. Statistical significance was determined using an independent t test. The underlying data for (A-F) can be found in Tab G in S1 Data. IAA, indole-3-acetic acid; RNAi, RNA interference; WT wild type.