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. 2019 Jan 29;11:9. doi: 10.3389/fnagi.2019.00009

Figure 3.

Figure 3

Proteasome capacity is diminished when HSP-110/70/J machinery is perturbed. (A) Scheme of the single-cell photo-conversion of UbG76V::Dendra2 in muscle and neuronal cells of C. elegans. (B,C) RNAi treated UbG76V::Dendra2 animals of muscle (B) or neuronal cells (C). UbG76V::Dendra2 protein was analyzed 3 (neurons) or 24 (muscle) hours post conversion at 561 nm. (B) Error bars represent the SD of a minimum of nine animals. (D,E) Immunoblotting of siRNA transfected HEK293 cells, expressing UbG76V-GFP. Cells treated with 20 μM MG-132 for 4 h served as control. The relative UbG76V-GFP levels are shown (E). Error bars represent the SD of three independent experiments. (F,G) Chymotrypsin-like proteasome activity of RNAi treated nematodes for 4 days. Shown are the relative activities (G). Error bars represent the SD of three independent experiments. (H,I) Relative protein levels of 20S α-subunits RNAi treated animals. (I) Error bars represent the SD of three independent experiments. (J,K) Chymotrypsin-like proteasome activity of siRNA treated HEK293 cells. The relative activities are depicted (K). The error bars represent the SD of three independent experiments. (L,M) Relative protein levels of 20S α-subunits of siRNA HEK293 cells. (M) Error bars represent the SD of a minimum of six independent experiments. (N,O) 20S α-subunit levels were determined of nematodes of 4, 6, 8 and 10 days of age and normalized to day 4 old animals (O). Error bars represent the SD of five independent experiments. (P) 20S α-subunits at day 4, 6, 8 and 10 of RNAi treated animals. Depicted are the averages of the relative 20S α-subunit levels from a minimum of three independent experiments.