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

Fig. 5. Role of KDM6B for the expression and function of ERβ and its response to KB9520 in A2780cis cells.

Fig. 5

a Representative RT-PCR and b Western blot analyses of KDM6B, SIRT1, and ESR2 expression in A2780cis cells transfected 48 h with non-specific or KDM6B-specific siRNAs. 18S rRNA and Tubulin were used as controls. c Immunoprecipitation of ERβ, from lysates of A2780cis cells 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 Immunoprecipitation of ERβ, from lysates of A2780cis cells transfected 48 h with non-specific or KDM6B-specific siRNAs and 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). e Representative Western blot analysis of PARP1 cleavage in A2780cis cells transfected with non-specific or 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. f Percentage of viable A2780cis cells transfected with non-specific or KDM6B-specific siRNAs after 24 h treatment with 40 μM of cisplatin alone or in combination with 10 nM of KB9520. Results are expressed as mean ± s.d. of three independent experiments. *p ≤ 0.05. g Representative RT-PCR, and h respective densitometry, of SIRT1 expression in A2780cis cells transfected with non-specific or KDM6B-specific siRNAs for 48 h and then treated or not with 10 nM of KB9520 for 2 h. 18S rRNA was used as housekeeping gene