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. 2019 Sep 27;33(12):13435–13449. doi: 10.1096/fj.201900722R

Figure 2.

Figure 2

PTEN down-regulation during hypoxia. A–C) Time course assay for PTEN expression levels in head and neck cancer cell lines cultured under hypoxic conditions and compared with normoxic controls Data represent means ± sem. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Note that all analyzed cancer cell lines present reduced levels of PTEN when cultured under hypoxia. D–F) Representative images from WSU-HN6, WSU-HN12, and WSU-HN13 cell lines cultured under the hypoxic condition for up to 18 h. Immunofluorescence staining for PTEN depicts down-regulation of the tumor suppressor starting after 6 h of hypoxia for WSU-HN6 and WSU-HN13 and after 12 h for WSU-HN12 compared with normoxic controls. pS6 staining was used as a readout for the activation of the PTEN/PI3K-signaling pathway. Arrows indicate high expression of pS6 after 12 h of hypoxia in all tested HNSCC cell lines. Scale bars, 100 μm.