Figure 5.
Anti-PRR intrabodies increase mHDx-1 turnover. A–C, ST14A cells were transfected with mHDx-1-SNAPtag 97Q (A, B) or wtHDx-1-SNAPtag 25Q (C) and intrabody at the optimal ratio for each intrabody. DAF green fluorescent SNAP substrate was added to cultures at 24 h after transfection. Some cultures were then fixed and stained with Toto-3 iodide nuclear marker, whereas others were incubated an additional 24 h to allow turnover of labeled HDx-1. Mean fluorescence intensity of cells at 24 h was compared with intensity at 48 h to determine the percentage of labeled HDx-1 remaining. VL12.3 has no effect on mHDx-1 or wtHDx-1 turnover, whereas MW7, Happ1, and Happ3 significantly increase the rate of mHDx-1 turnover. n = 4; *p < 0.01.