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. 2019 Nov 18;130(1):83–93. doi: 10.1172/JCI127429

Figure 5. CNO washout causes reversal of the PA phenotype in AS+/Cre hM3Dq mice.

Figure 5

(A) Experimental protocol. Female 20- to 22-week-old AS+/Cre hM3Dq mice were treated with a HS diet for 3 days. All mice were then administered HS plus CNO for 7 days. One group was sacrificed on day 7 (HS + CNO), whereas the other group underwent a washout (WO) protocol for an additional 10 days with a HS diet but without CNO (HS + CNO WO). (B) qPCR analysis of whole adrenal Cyp11b2 mRNA. For qPCR analysis, HS plus vehicle treatment, n = 7; for HS plus CNO treatment, n = 5; and for HS plus CNO washout, n = 7. (C) LC-MS/MS measurement of plasma aldosterone levels. For steroids, HS plus vehicle treatment, n = 8; for HS plus CNO treatment, n = 6; and for HS plus CNO washout, n = 7. (D) Immunofluorescence staining for Cyp11b2 (red) and Cyp11b1 (green). Scale bars: 50 μm. For B and C, the previously described controls treated with HS plus vehicle (Figure 4) were used as a comparison for the HS plus CNO and HS plus CNO washout groups. Data in the dot plots represent the mean ± SEM. ***P < 0.001, by 1-way ANOVA with Bonferroni’s correction.