Fig 1: UbVQ40V/T66K/V70F displays selective inhibition of UCHL3. The percent activity of each enzyme normalized to 100% activity for the DMSO control as a function of UbV concentration are plotted for UCHL3 (blue) and UCHL1 (red).
Fig 2: Time-dependent Western blot for Ub-ABPs. HA-WT-Ub-PRG and HA-UbVT66K/V70F-PRG, 5 µM, were both dosed into HEK293 lysates (0.5 mg/mL). Samples were tested at the indicated time points. (Top) Western blot for UCHL3 indicating the free-UCHL3 and Ub-UCHL3 complex gel shifted. (Bottom) Western blot for HA (green fluorescence) and UCHL3 (red fluorescence).
Fig 3: UCHL3 and NEDD8 gene expression levels in each group. Relative gene expression levels in different (A) A375 and (B) SK-MEL-2 cell groups. ***P<0.001 vs. normal control group. UCHL3, ubiquitin C-terminal hydrolase-L3; NEDD8, neural precursor cell-expressed developmentally downregulated protein 8; NC, negative control; si, small interfering.
Fig 4: Detection of cell apoptotic rates in each group. Apoptotic rates in (A) A375 and (B) SK-MEL-2 cell groups. ***P<0.001 vs. normal control group. UCHL3, ubiquitin C-terminal hydrolase-L3; NEDD8, neural precursor cell-expressed developmentally downregulated protein 8; NC, negative control; si, small interfering.
Fig 5: Effect of UCHL3 knockdown on relevant protein expression and NEDD8 ubiquitination. (A) NEDD8 and LC3 protein expression levels were detected via western blotting. (B) NEDD8 ubiquitination. ***P<0.001 vs. normal control group. UCHL3, ubiquitin C-terminal hydrolase-L3; NEDD8, neural precursor cell-expressed developmentally downregulated protein 8; NC, negative control; si, small interfering.
Supplier Page from Abcam for Anti-UCHL3 antibody [EP5331]