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In vitro analysis of the HOL reporter response to HIF-1 activation.

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posted on 10.11.2011, 01:12 by Tetsuya Kadonosono, Takahiro Kuchimaru, Shuichi Yamada, Yumi Takahashi, Atsushi Murakami, Taeko Tani, Hitomi Watanabe, Tomoharu Tanaka, Kiichi Hirota, Masahiro Inoue, Tetsuya Tsukamoto, Takeshi Toyoda, Koji Urano, Kazuhiko Machida, Tomoo Eto, Tomoyuki Ogura, Hideki Tsutsumi, Mamoru Ito, Masahiro Hiraoka, Gen Kondoh, Shinae Kizaka-Kondoh

(A) MEF cells from FVB/HOL (Tg) and FVB/N (non-Tg) mice were cultured under hypoxic conditions (1% O2) for the indicated time, and luciferase activity was measured. The experiments were performed in triplicate, and the mean luciferase activity ± SD is shown in the graph. *P<0.05, **P<0.005. (B) FVB/HOL mice were peritoneally injected with PG, and 2 h later RNA was isolated from the indicated tissues. RT-PCR analysis was performed with the isolated RNA. Glyceraldehyde-3-phosphate dehydrogenase was used as an internal control. +, indicates a transgene used as a template; −, indicates no template. The experiment was performed in triplicates, and the representative data are shown. (C) FVB/HOL (Tg) and FVB/N (non-Tg) mice were peritoneally injected with PG, and 2 h later the mice were injected with luciferin through the tail vain; 2 min later, the tissues were isolated and observed by ex vivo bioluminescent imaging. a, liver; b, intestine; c, lung; d, uterus; e, kidney; f, spleen and pancreas; g, heart; h, thymus; i, bladder.

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