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. Author manuscript; available in PMC: 2018 Aug 13.
Published in final edited form as: Oncogene. 2018 May 2;37(32):4385–4397. doi: 10.1038/s41388-018-0278-0

Figure 3. Radiation induces BMP-mediated SIRT1 expression.

Figure 3

a. Fold change of SIRT1 mRNA in benign prostate and prostate cancer patients, obtained from R2-Genomics analysis is expressed (n = 95). b. SIRT1 mRNA expression was measured in 22Rv1 under serum free conditions with 50 ng/ml BMP4, in the context of increasing doses of TRC105 (0.05, 0.1, 0.5, 1, 5, or 10 µg/ml) or with 50 ng/ml noggin. SIRT1 mRNA expression was normalized to GAPDH and to serum treated control. c. Western blot for phosphorylated Smad1/5, SIRT1 and β-actin expression was measured in 22Rv1 cells following serum starvation and treatment with 50 ng/ml BMP4. Densitometric quantitation is indicated under each band. d. SIRT1 and p21 protein expression was measured 72 hours after irradiating (4 Gy) 22Rv1 and PC3 cells in the presence of IgG or TRC105 prior to irradiation. e. γ-H2AX (green) was immunolocalized in PC3 cells following treatment with IgG, TRC105, and 4 Gy irradation at 4 hours. Cells were nuclear counterstained with DAPI (blue). Foci per nuclei were quantified (n = 100). (****p<0.0001 as compared to control). f. Clonogenic survival assay was performed on p53 null PC3 cells at indicated doses of radiation. No significant (NS) radiation sensitization was had with TRC105 compared to IgG control. mRNA expression are reported as a mean +/− S.D. (**p < 0.01, ***p < 0.001, ****p < 0.0001, compared to control).