Figure 1

CXCR7 expression in prostate-cancer cells. (A) Western blot of 1, 2, and 4 μg of LNCaP total lysate with pAbCXCR7 antibody in the presence of the non-competitive AR peptide (a.a. 299–315, left panel) or the CXCR7 blocking peptide (a.a. 348–362, right panel) as detailed in Materials and Methods section. The CXCR7 bands are indicated by arrowheads. (B)AR, CXCR7, CXCR4, and PSA gene expressions in LNCaP cells transfected with AR, CXCR7, or scrambled control siRNA. RNA was isolated 72 hrs post-transfection and measured by qPCR. Student’s t-test was used to calculate significant differences (*p ≤ 0.05, n = 3) between control and experimental cells. (C) Western blot (left panel) of whole cell lysates from LNCaP cells transfected with control or two experimentally-validated CXCR7 siRNAs (CXCR7 #1 or #2) for 72 hrs using antibodies to pAbCXCR7 and GAPDH. Silver-stained gel demonstrated equal protein loading across samples (right panel). The densitometry values were labeled below the blot and normalized to the control transfected cells loaded with the same amount of total proteins. (D) Light microscopy of LNCaP cells transfected with control or CXCR7 siRNA for 72 hrs.