FIGURE 5.

The TMCD is required for the translation-independent ER localization of ALPP. COS-7 cells were transfected with plasmid containing either ALPP or ALPP-ΔTMD and allowed to express mRNA for 18–24 h. The cells were treated with DMSO (Ctrl) or HHT for 30 min and then extracted with digitonin. The cells were fixed and stained for mRNA using a specific FISH probe against the ALPP ORF. A, representative examples. B, quantification of the fluorescence intensities of mRNAs in the ER and nucleus. All data were normalized to the ER staining intensity in the control treated group for each construct. Each bar represents the average ± S.E. (error bars) of three independent experiments (each experiment consisting of n>30 cells). Scale bar, 20 μm.