ER stress is less severe in Ad-DHCR24-infected N2A cells.

<p>N2A cells were infected with the indicated adenoviruses for 2 days, and then exposed to TM for various lengths of time. Whole cell lysates (50 µg/lane) were subjected to western blot analysis using anti-Bip, anti-CHOP, anti-myc or anti-actin antibody. The proteins were visualized using the ECL method. Representative results are shown in panel A. The expression levels of Bip and CHOP were normalized against actin level, and are expressed as percentage of the maximal level in the cells infected with Ad-LacZ. n = 3, Mean ± S.D., *: p<0.05 versus the levels in cells infected with Ad-LacZ. Cells were cultured on cover slips and infected with adenoviruses for 2 days, and then exposed to TM for the indicated time points, followed by immunocytochemical analysis. Cells were probed with mouse anti-CHOP antibodies, followed by anti-mouse IgG antibody conjugated to Alexa fluor-488. Nuclei were stained with PI. Images were obtained by confocal laser microscopy. Scale bar, 50 µm (Panel C). Scale bar, 5 µm (Panel D). The relative fluorescence signal intensities of Panel C were represented as percentage relative to that at time 0. Values are expressed as mean ± S.D. (n = 3) *p<0.05 versus Ad-LacZ. Similar results were obtained from three separate experiments.