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. Author manuscript; available in PMC: 2012 Aug 1.
Published in final edited form as: Prostate. 2011 Jan 12;71(11):1198–1209. doi: 10.1002/pros.21335

Figure 2. Nodal Expression in Prostate Cancer Cell Lines.

Figure 2

(A) Real-time PCR of Nodal expression in prostate cancer cell lines normalized to H9 hESCs (n=3). (B) Western blots of lysates from prostate cancer cell lines immunoprecipitated using rabbit or goat anti-Nodal antibodies. LNCaP cells stably transfected with Nodal were used as a positive control for the Western blot (Nodal Input) and IP (Nodal IP). All blots were probed with a mouse anti-Nodal antibody. Nodal was detected as a single ~42 kDa band. The unbound fractions of IP samples were probed for GAPDH to compare initial protein concentrations. (C) RT-PCR showing that Cripto-1 was expressed in all prostate cell lines. H9 hESC was used as a positive control and GAPDH as the house-keeping gene. (D) Western blots of prostate cancer and MEL1 hESC lysates where Cripto-1 was detected as two bands of approximately 17 and 25 kDa. GAPDH was used as a loading control. (E) RT-PCR showing negligible Lefty and Cerberus expression in prostate cancer cells compared to H9 hESCs. Low levels of TMEFF1 were expressed in 22Rv1 and DU145 cells. LN = LNCaP, 22 = 22Rv1, DU = DU145, MD = MDAPCa2b, SC = H9 or MEL1 hESCs, RTase = reverse transcriptase, and NTC = no template control.