Figure 6
Optical Pulse Labeling reveals no effect of UBQLN2 on α-syn clearance in neurons. (A) Schematic of Optical Pulse labeling experiment. Rat primary cortical neurons were transfected with hSyn-Dendra2 and UBQLN2-iRFP or iRFP. Dendra2 was photoconverted 24 h post-transfection with 405 nm light, imaged immediately and every 24 h thereafter for 3 days. Scale bar 25 μm. (B) Optical pulse labeling reveals no change in red fluorescence intensity over time in neurons transfected with UBQLN2 (n = 213) compared to control (n = 182; p = 0.6). (C) Representative images of optical pulse labeling of hSyn-Dendra2 in rat primary cortical neurons transfected with hSyn-Dendra2 and UBQLN2-iRFP or iRFP. (D) Optical pulse labeling in primary rate neurons reveals no change in half-life of hSyn-Dendra2 upon transfection with UBQLN2 vs control (iRFP; p = 0.28). hSyn-Dendra2 half-life was determined by measuring the loss of photoconverted hSyn-Dendra2 (RFP) signal over time for each cell. (E) Density plot of hSyn-Dendra2 half-life measurements from individual neurons transfected with control (n = 182) or UBQLN2 (n = 213). Values were pooled from eight wells per condition, from each of two biological replicates and analyzed by the Kruskal–Wallis test.
