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. Author manuscript; available in PMC: 2019 Jun 1.
Published in final edited form as: Prostate. 2018 Mar 11;78(8):595–606. doi: 10.1002/pros.23504

Figure 2. Confirmation that HS-5 BMSC conditioned medium modulates select genes identified in the 1,046 gene expression profile.

Figure 2

(A, B, C) RT-QPCR and (C) western blot analysis was performed for the AR+ LNCaP PCa cell line or the AR− PC3 PCa cell line treated for 3 days with PCa control conditioned medium (Control CM), HS-5 BMSC CM, or HS-27a BMSC CM. (A) PC3 cells have little or no detectable AR, PSA, NKX3.1, or TMPRSS2 mRNA relative to LNCaP cells. (B) HS-5 CM can repress AR, PSA, NKX3.1, and TMPRSS2 mRNA in LNCaP cells. (C) HS-5 CM can induce p62, ELF3, SOX9, CD24 and CD44 mRNA LNCaP cells. PC3 cells have high basal p62, ELF3, SOX9, CD24, and CD44 mRNA relative to LNCaP cells and show no significant induction response to HS-5 CM for these genes. (D) HS-5 CM can induce p62 protein and downregulate AR and NKX3.1 protein in LNCaP cells. PC3 cells do not accumulate AR or NKX3.1 protein and show no detectable p62 induction response to HS-5 CM. HS-5 CM induces SOD2 mRNA and protein in both LNCaP and PC3 cells and is used as a treatment efficacy control. n = 4 biological replicates (Bio-Rep); error bars = +/−STDEV; p-value = * ≤ 0.05, ** ≤ 0.005, ***≤ 0.0005. mRNA fold change is normalized to LNCaP vehicle control. β-actin is a western blot loading control.