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. Author manuscript; available in PMC: 2016 Jan 26.
Published in final edited form as: N Engl J Med. 2013 Nov 28;369(22):2105–2114. doi: 10.1056/NEJMoa1304603

Figure 4. Analysis of the Role of ARMC5.

Figure 4

Panels A and B show the effects of ARMC5 inactivation by siRNA on the expression of ARMC5, NR5A1, NR0B1, MC2R, CYP17A1, and CYP21A2 and on basal and forskolin-stimulated cortisol synthesis in adrenocortical H295R cells. NS denotes not significant. Panel C shows immunofluorescence staining of transfected H295R cells with different ARMC5-FLAG constructs (a nonmutated construct and two missense mutants: p.R898W and p.L548P). After 6 hours, the cells expressing the ARMC5 constructs had normal morphologic features. After 14 hours, the cells expressing the nonmutant ARMC5 construct (white arrows) were apoptotic (coexpression of the cleaved caspase 3 [green stain] and altered cell morphology with condensed nuclei), and the cells expressing the missense ARMC5 constructs (p.R898W and p.L548P) were not apoptotic. Red staining shows FLAG antibodies (ARMC5 constructs), and blue staining shows 4′,6-diamidino-2-phenylindole (DAPI) (cell nuclei).