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Fig. 1 | Journal of Experimental & Clinical Cancer Research

Fig. 1

From: CRISPR/Cas9 targeting of GPRC6A suppresses prostate cancer tumorigenesis in a human xenograft model

Fig. 1

Expression and function of GPRC6A. a RT-PCR analysis of GPRC6A message expression in human prostate cancer cell lines, DU145, PC-3, LNCap and 22Rv1. b Comparison of nucleotides and amino acid sequences of GPRC6A 3rd IL from human GPRC6A reference (NM_148963) sequence showing the K..Y insertion/deletion, and the presence of the ancestral RKLP sequence in the human rs386705086 GPRC6A SNP, mouse GPRC6A (NM_153071) and bovine GPRC6A (XM_010808417) sequences. c Data from www.1000genomes.org showing distribution of the RKLP allele in human grouped by geographical area. Each population group contains a minimum of 4 and maximum of 7 subpopulations. The ancestral RKLP polymorphism is overrepresented in people of African descent. d Comparison of GPRC6A cell-surface distribution in HEK-293 cells transfected with mouse GPRC6A (upper panel) and “humanized” GPRC6A, where the “K..Y” sequences replaces the RKLP sequence in the mouse cDNA (lower panel). Transfected cells were fixed and stained with anti-myc antibodies and imaged on a Zeiss Axiolmager II microscope. Bars are 10 μm. e Immunoblot analysis of total and membrane-associated GPRC6A from PC-3 cells transfected with the mouse GPRC6A cDNA. Consistent with membrane expression in d, GPRC6A was enriched in the membrane fraction. f mGPRC6A-WT (left panel) and mGPRC6A-KY mutant (right panel) activity as assessed by phosphorylation of mTOR, p70S6K (S6K), and ERK. * Significant difference from control group and sgRNA3 group at p < 0.05 (n ≥ 4). g Comparison of mGPRC6A and hGPRC6A response to the ligands, 200 nM testosterone, 160 ng/ml osteocalcin and 10 mM L-Arg in HEK-293 cells transfected with mouse GPRC6A or human GPRC6A cDNAs

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