Figure S1.

Characterization of readthrough quality control machinery, related to Figure 1

(A) Readthrough into 3′ UTR induces expression of sHSPs. Volcano plot representation of label-free proteome analysis of YFP-UTR and YFP-STOP expressing nematodes. sHSPs are highlighted in red. See Table S1K.

(B) Relative mRNA levels (qPCR analysis) of hsp-16 family members and hsp-70 (C12C8.1) in animals (day 0) expressing YFP-UTR. Wild-type (WT) nematodes (untreated or exposed to heat stress [HS]) and animals expressing YFP-STOP were analyzed as controls. HS was performed for 60 min at 34°C. Error bars represent mean ± SEM (n = 3).

(C) Expression of readthrough reporter protein in C. elegans muscle cells in the presence of a HSP-16.1-RFP reporter. Representative fluorescence microscopy images of animals expressing YFP-UTR are shown.

(D) YFP-UTR undergoes proteasomal degradation. Representative fluorescence microscopy images of wild-type, Δrnf-126, and Δrpn-10 worms expressing YFP-UTR. (Exposure 200 ms, 100× magnification.)

(E) Hydrophobicity analysis of YFP-UTR using Kyte Doolittle scores (KDSs) as metric. AA, amino acid.