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. 2009 Sep 16;106(39):16805–16810. doi: 10.1073/pnas.0904606106

Fig. 4.

Fig. 4.

HIF-2α knockdown induces early sympathetic differentiation in TICs. (A) HIF-2α knockdown at the protein level assessed by Western blot analysis. Dividing lines indicate that two areas of the same film have been merged. (B) HIF2A knockdown at the mRNA level assessed by qPCR. (C) Down-regulation of VEGF mRNA upon HIF-2α knockdown. (D) icNotch-1 is not affected by HIF-2α knockdown, as measured by Western blot analysis of nuclear extracts from shC and shHIF2A TICs. (E–G) Down-regulation of Notch pathway components upon HIF-2α knockdown assessed by qPCR. (H–J) Up-regulation of sympathetic neuronal differentiation markers at the mRNA level upon HIF-2α knockdown assessed by qPCR. (K) Up-regulation of chromogranin A protein by HIF-2α knockdown as shown by immunohistochemistry. (L–N) Up-regulation of sympathetic neuronal differentiation markers at the mRNA level upon rapamycin treatment in TICs, as determined by qPCR. Error bars represent SD. (O–Q) Neuronal differentiation markers are not induced by rapamycin in shHIF2A cells, as determined by qPCR. The qPCR data are mean values of three independent experiments performed in triplicate or representative data of three independent experiments performed in triplicate (L–Q). Error bars represent SEM. Immunohistochemistry and Western blot analysis data were repeated at least three times.