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. 2022 Feb 9;298(3):101707. doi: 10.1016/j.jbc.2022.101707

Figure 6.

Figure 6

ATP1A1 knockdown enhances cardiac glycoside-mediated inhibition of STAT1(Y701) phosphorylation.A, representative levels of pSTAT1(Y701) and total STAT1 are shown for the MDA-MB-231 ouabain and digoxin experiments in Figure 4. For loading controls, see Figure 4, D and E. B, representative levels of pSTAT1(Y701) and total STAT1 are shown for the A549 ouabain and digoxin experiments in Figure 5. For loading controls, see Figure 5, C and D. The dashed vertical lines indicate one lane (ladder) that has been spliced out. C, quantification of pSTAT1(Y701) in western blots of NTC or ATP1A1 siRNA transfected TNF/IFN-ɣ (24 h) stimulated MDA-MB-231 cells treated with 100 nM ouabain. ∗p < 0.05, ∗∗∗p < 0.001, Two-factor ANOVA, Tukey’s multiple comparisons. D, STAT1 activation time-course experiment in MDA-MB-231 cells treated with 100 nM ouabain or 0.05% DMSO and stimulated for 30 min, 2 h, 8 h, or 24 h with TNF/IFN-γ. Mean intensities ±standard deviation, measured by densitometry of blots from three independent time-course experiments, are listed under the pSTAT1(Y701) long and total STAT1 panels.