Picture from Darom A et al. (2010) Mol Biol Cell "RNF-121 is an endoplasmic reticulum-membrane E3 ubiquitin ligase involved in ...."

Figure 1. Topology prediction, expression pattern, and in vitro E3 ligase activity of RNF-121. (A) Schematic representation of the predicted membrane topology of RNF-121. Six TMDs are predicted, the RING finger domain faces the cytosol. The amino acid sequence of the RING finger domain is shown. The cysteine and histidine residues of the RING finger domain are shown in red. The predicted E2 binding site (V224) is labeled in yellow (B and C) RNF-121::GFP is localized to the ER in body wall muscles (mu), hypodermis (hyp), and seam cells (SE). Bar, 10 um. (D) RNF-121::GFP colocalizes with the mRFP::SP12 reporter. Bar, 5 um. (E and F) RNF-121::GFP is expressed in the somatic gonad and vulva. Lateral view of midL4 animal. Expression is detected in the vulval cells (vul), spermathecal cells (sp), uterine cells (ut), and the distal tip cell (DTC). Bar, 10 um. (G) RNF-121::GFP colocalizes with the mRFP::mans reporter. Bar, 5 um. (H) In vitro self-ubiquitination assays were performed using wild-type GST-RNF-121RING or RING mutant forms of bacterially expressed and purified protein. In lane 4 (-), the reaction includes E1, E2, Ub, and ATP without the E3 ligase. GST-RNF-5 served as a positive control. Arrow indicates the E1 enzyme. Arrowhead indicates the 5% stacking gel-10% running gel boundary. High-molecular-weight smears generated by autoubiquitination of the indicated E3 are labeled with parenthesis. Coomassie-stained gel of the purified glutathione transferase (GST)-tagged expressed proteins is shown (bottom).

Picture from Cornils A et al. (2016) PLoS Genet "Structural and Functional Recovery of Sensory Cilia in C. elegans IFT Mutants ...."

A) Expression of osm-5::gfp under the srg-47 promoter sequences restores ASI cilialength in osm-5(ok451) but not osm-6(p811) mutants. White and yellow arrowheadsindicate cilia base and cilia axoneme, respectively. Magnified images of cilia are shownin the insets. Anterior is at top. Scale bar: 10 um; insets 5 um.B) Ratio of fluorescence levels of Ub-G76V::GFP to TagRFP in the ASI soma of 1d and4d old animals of the indicated genotypes grown at 20 degrees C (left) and 25 degrees C (right). Bothtransgenes were expressed under srg-47 promoter sequences. *** indicates different from1d of the same genotype at P<0.001 (Wilcoxon Mann-Whitney U test). n>45 each; 3independent experiments.C) Large and small aggregates of SOD1(G85R)::YFP protein in body wall muscle andASI soma in 1d and 4d old wild-type and osm-6(p811) mutants. Expression was drivenunder the unc-54 (muscle) and srg-47 promoters (ASI). Large and smallSOD1(G85R)::YFP aggregates were defined as puncta that were >3 um and <1 um indiameter, respectively. Scale bar: 10 um.(D) Average number of small and medium sized aggregates of SOD1(G85R)::YFP in theASI soma of 1d and 4d old animals of the indicated genotypes. Expression in ASI wasdriven under the srg-47 promoter. Animals also expressed SOD1(G85R)::YFP undermuscle specific unc-54 regulatory sequences (see S5C Fig). Small and mediumSOD1(G85R)::YFP aggregates were defined as puncta that were <1 um and between 1-3 um in diameter, respectively. ## and #indicate different from 1d within a genotype atP<0.005 and 0.05, respectively (Wilcoxon Mann-Whitney U test). n>30 each; 3independent experiments.E) Quantification of srg-47p::TagRFP levels in ASI neurons of wild-type and osm6(p811) animals of the indicated ages. n>45 neurons each; 3 independent experiments.AU-arbitrary fluorescence units.

Picture from Cornils A et al. (2016) PLoS Genet "Structural and Functional Recovery of Sensory Cilia in C. elegans IFT Mutants ...."

S5 Fig. Proteostasis is improved in ciliated sensory neurons in agedanimals.A) Expression of osm-5::gfp under the srg-47 promoter sequences restores ASI cilialength in osm-5(ok451) but not osm-6(p811) mutants. White and yellow arrowheadsindicate cilia base and cilia axoneme, respectively. Magnified images of cilia are shownin the insets. Anterior is at top. Scale bar: 10 um; insets-5 um.B) Ratio of fluorescence levels of Ub-G76V::GFP to TagRFP in the ASI soma of 1d and4d old animals of the indicated genotypes grown at 20 degrees C (left) and 25 degrees C (right). Bothtransgenes were expressed under srg-47 promoter sequences. *** indicates different from1d of the same genotype at P<0.001 (Wilcoxon Mann-Whitney U test). n>45 each; 3independent experiments.C) Large and small aggregates of SOD1(G85R)::YFP protein in body wall muscle andASI soma in 1d and 4d old wild-type and osm-6(p811) mutants. Expression was drivenunder the unc-54 (muscle) and srg-47 promoters (ASI). Large and smallSOD1(G85R)::YFP aggregates were defined as puncta that were >3 um and <1 um indiameter, respectively. Scale bar: 10 um.(D) Average number of small and medium sized aggregates of SOD1(G85R)::YFP in theASI soma of 1d and 4d old animals of the indicated genotypes. Expression in ASI wasdriven under the srg-47 promoter. Animals also expressed SOD1(G85R)::YFP undermuscle specific unc-54 regulatory sequences (see S5C Fig). Small and mediumSOD1(G85R)::YFP aggregates were defined as puncta that were <1 um and between 1-3um in diameter, respectively. ## and #indicate different from 1d within a genotype atP<0.005 and 0.05, respectively (Wilcoxon Mann-Whitney U test). n>30 each; 3independent experiments.E) Quantification of srg-47p::TagRFP levels in ASI neurons of wild-type and osm6(p811) animals of the indicated ages. n>45 neurons each; 3 independent experiments.AU-arbitrary fluorescence units.