Figure 1 | Scientific Reports

Figure 1

From: Hypoxia, acidification and oxidative stress in cells cultured at large distances from an oxygen source

Figure 1

(A) Schematics of the cell chamber used in the coverslip-induced hypoxia variant. An acrylic coverslip with a central hole is placed on top of the well of a 35 mm glass bottom dish. The central hole is the only oxygen source, and cells inside the well are at varying distances from it. (B) Representative images of pimonidazole adduct detection (Hypoxyprobe), shown here in HaCaT E5/E6/E7 HPV-18 cells cultured under coverslips for 24 h. Images from the central zone or the periphery of the cell chamber are shown. (C) Quantification of O2 levels in HaCaT parental and HaCaT E5/E6/E7 HPV-18 cells in normoxia and with a coverslip. (D) BioTracker™ 520 Green Hypoxia Dye signal in control cells, or in cells under coverslips for 24 h, either in the central zone or the periphery of the cell chamber. (E) Quantification of hypoxia signal through BioTracker Green Hypoxia Dye in HaCaT E5/E6/E7–18 and parental HaCaT cells. (*) P < 0.05, (**) P < 0.01, (***) P < 0.001. Scale bar: 20 μm.

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