Fig. 2

HDAC6 inhibition results in the accumulation of both cytoplasmic and cell surface Cx32. a TubA treatment stabilizes Cx32 levels as determined by increases in levels of biotinylated Cx32 at 1.5 h after labeling. N2a cell were transfected with Cx32-Myc/6xHis and treated with TubA or vehicle for 3 h prior to biotinylation of cell surface proteins. Following cell surface biotinylation, cultures were harvested and processed for total lysate (top) and biotinylated Cx32-Myc/6xHis (bottom) by Western blot, as described in Methods. b Densitometric quantification of Western blots shows a significant reduction in biotinylated Cx32/total Cx32 at T0 in TubA-treated samples as compared to vehicle-treated cells. c A significant increase in biotinylated Cx32/total Cx32 remained at T1.5 compared to vehicle-treated (N = 3 independent experiments, Student’s T-test *p < 0.05). d Immunostaining and confocal microscopy cell surface labeling of Cx32 confirmed TubA treatment increases Cx32 accumulation in cell surface and cytoplasmic compartments (see also Additional file 5: Fig. S1). Images shown were obtained at 63X magnification and scale bars = 25 μm. Arrows indicate points of cell-cell contact