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. 2016 Jul 13;8(23):37041–37060. doi: 10.18632/oncotarget.10556

Figure 3. Effects of PML-RAR, RARα2 and RARα1 knock-down on the growth and differentiation of NB4 cells.

Figure 3

A. The indicated NB4 cell populations stably infected with shRNAs targeting PML-RAR (PMRsh-NB4), RARα1 (RA1sh-NB4), RARα2 (RA2sh-NB4) or the control scramble shRNA (SCRsh-NB4) were grown in the presence of vehicle (DMSO) or ATRA (1 μM) for the indicated amount of time. The number of viable cells determined after staining with trypan blue is indicated. Each point is the mean±S.D. of three replicate cultures. ** = Significantly different relative to the corresponding SCRsh-NB4 time point (p<0.01 after Student's t-test); * = Significantly different relative to the corresponding SCRsh-NB4 time point (p<0.05 after Student's t-test). B. The indicated NB4 cell populations [see (A)] were grown in the presence of vehicle (DMSO) or ATRA (1 μM) for 72 hours. Cells were subjected to FACS analysis for the indicated markers. The column graphs on the left indicate the percentage of CD11b-, CD11c- and CD38-positive cells. The graphs on the right indicate the MAF (mean-associated-fluorescence) values determined. The results are representative of two independent experiments. C. The indicated NB4 cell populations were treated as in (B) for 48 hours. Cell extracts were subjected to Western blot analysis for the indicated proteins. Actin is used as a loading control. The results shown in the upper and lower panels were obtained in separate experiments. Each line shows cropped lanes of the same gel, hence the results can be compared across the lanes, as they were obtained with the same exposure time. The calculated molecular weight (M.W.) of each protein is indicated on the left. The results are representative of at least two independent experiments.