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. 2018 Feb 9;7(2):15. doi: 10.1038/s41389-018-0027-9

Fig. 6. Role of SIRT1 for the expression and function of ERβ and its response to KB9520 in A2780cis cells.

Fig. 6

a Representative RT-PCR of SIRT1, KDM6B, and ESR2 expression and b Western blot analyses of ERβ expression in A2780cis cells transfected with non-specific or SIRT1 and KDM6B-specific siRNAs. 18S rRNA and Tubulin were used as controls. c Immunoprecipitation of ERβ, from lysates of A2780cis cells transfected for 48 h with non-specific or SIRT1-specific siRNAs and then treated or not with 10 nM of KB9520, 2 h; lysine acetylation and co-immunoprecipitated proteins were detected by Western blot analyses using the respective antibodies (AcLys, ERβ, and p300). d Representative Western blot and e densitometric analysis of PARP1 cleavage in A2780cis cells transfected with non-specific or SIRT1 and KDM6B-specific siRNAs after 24 h treatment with 40 μM of cisplatin alone or in combination with 10 nM of KB9520. Tubulin was used as loading control. *p ≤ 0.05. f Immunoprecipitation of p300, from lysates of A2780cis treated or not 1 h with 10 nM of KB9520; co-immunoprecipitated proteins were detected by Western blot analyses using the respective antibodies (p300, SIRT1, and ERβ)