Figure 4: The Pacer RH domain basic triad controls subcellular localization [FINAL]

Figure 4. The Pacer RH domain basic triad controls subcellular localization. (A) Fluorescent bead binding experiment performed as in Figure 2D, and quantified as in Figure 3D. Mutants of MBP-Pacer RH with the listed mutant were used in these experiments. (B) Quantification of panel A, conducted as in Figure 3E. Each experiment consists of ~20 different beads measured and averaged. This was repeated 3 times, and the mean of these biological replicates is plotted. Error bars indicate standard deviation of separate experiments. (C) Colocalization of mCherry- wild-type Pacer and mCherry-Mutant (K534N, R623T) Pacer. Cells were transfected with the listed Pacer construct, and depolarized for 1 H using CCCP. Immunofluorescence was performed using the pS72 specific antibody of RAB7A, and the Alexa 488 secondary antibody was used to mark pRAB7A. Insets are digitally expanded images, 2.8 μm wide. (D) Linescans indicated in panel C, calculated as in Figure 3B. (E) Colocalization of WT and Mutant Pacer with pRAB7A, calculated as in Figure 3C.