Grb2 helps in Htt clearance via lyso-autophagy pathway and also in autophagosome fusion with Lysosome

<p>A. Western blot analysis of expression of Grb2, pERK1/2, total ERK, LC3 and beta actin in STHdhQ^7/7, STHdhQ^7/7 transfected with Grb2-Dsred, STHdhQ^7/7 transfected with Dsred, STHdhQ^7/7 Grb2si, STHdhQ^7/7 Scrmbsi, STHdhQ^111/111 transfected with Grb2-Dsred, STHdhQ^111/111 transfected with Dsred, STHdhQ^{111 /111}Grb2si and STHdhQ^111/111 Scrmbsi cells. B. Bar diagram of pERK1/2 levels of three experiments (n=3) as shown in panel A. normalized to total ERK1/2 levels. C. Bar diagram of LC3 II levels of three experiments (n=3) as shown in panel A normalized to beta actin levels. D. Representative confocal images of STHdhQ^7/7, STHdhQ^111/111, STHdhQ^{111 /111}Grb2si, STHdhQ^111/111 Scrmbsi cells transfected with dual tagged GFP-LC3-mCherry at 24hrs, 48hrs, 72hrs and 96hrs timepoints. E. Bar diagram of ratio of number of green/red to free red vesicles in each cell of D (n≥20). F. Representative western blot showing LC3 and beta actin levels of STHdhQ^7/7, STHdhQ^111/111 and STHdhQ^{111 /111}Grb2si cells treated or untreated with 100nM bafilomycinA1, G. Bar diagram of three independent (n=3) experiments of F. of LC3II levels normalized to beta actin levels.