Figure 1

Lentivirus-mediated shRNA efficiently inhibited the expression of HIF‑1α in head and neck squamous carcinoma Cal27 and SCC2 cells. A. Fluorescence microscopic picture of the expressed reporter Turbo-RFP protein as a marker of successful transfection and doxycycline indelibility. Lentivirus doxycycline-inducible pTRIPZ-shRNAmir constructs directed against HIF‑1α used to transfect Cal27 and SCC25 HNSCC cells. The construct also contains a doxycycline-inducible TurboRFP as a reporter gene co-expressed with the shRNA sequence. B. Immunoblot with anti- HIF‑1α showed that doxycycline-induced HIF-shRNAmir reduced the HIF‑1α protein level by 90% relative to the untreated cells. Cobalt chloride at 2mM was added for 16h to induce hypoxia in the cell and induce HIF‑1α.