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. 2014 Sep 2;22(11):1936–1948. doi: 10.1038/mt.2014.134

Figure 4.

Figure 4

Intra-cranial and s.c. tumor phenotypes are distinct. (a) cDNA from B16-ova cells freshly resected from s.c. tumors (SCT) (Lanes 1–5), B16-ova cells freshly resected from three pooled i.c. tumors (ICT) (Lanes 6–10), B16-ova cells resected from three pooled i.c. tumors and maintained in culture for 3 weeks (Lanes 11–15) or long-term in vitro cultured B16-ova cells (Lanes 16–20) was screened by qRT-PCR for expression of HIF-2α (Lanes 1, 6, 11, 16), C-MYC (Lanes 2, 7, 12, 17), TYRP-1 (Lanes 3, 8, 13, 18), N-RAS (Lanes 4, 9, 14, 19) and CYT-C (Lanes 5, 10, 15, 20). The difference in cycle threshold (CT) for expression of the control GAPDH gene and the target gene (CT(GAPDH)-CT(Target gene)) is shown. Results are representative of three separate experiments. (b) HIF-2α protein expression from B16-ova cells freshly resected from s.c. tumors (Lane 1), B16-ova cells freshly resected from three pooled i.c. tumors (Lane 2), B16-ova cells resected from three pooled i.c. tumors and maintained in culture for 3 weeks (Lane 3) or long-term in vitro cultured B16-ova cells (Lane 4) was measured by ELISA with samples standardized for equal protein loading. (c) The experiment of b was repeated using glioma GL261 cells freshly resected from i.c. tumors (Lane 1) or long-term in vitro cultured GL261 cells (Lane 2); prostate TC2 cells freshly resected from s.c. tumors (Lane 3) or long-term in vitro cultured TC2 cells (Lane 4). (d) N-RAS or (e) C-MYC protein expression from long-term in vitro cultured B16-ova (Lane 1) or GL261 (Lane 4) cells, B16-ova (2) or GL261 (5) cells freshly resected from i.c. tumors, or B16-ova (3) or GL261 (6) cells freshly resected from s.c. tumors, was measured by Western Blot with samples standardized for equal protein loading using β-actin.