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. 2022 Mar 1;12:842418. doi: 10.3389/fonc.2022.842418

Figure 2.

Figure 2

Increased GRP78 expression on the surface of irradiated HNSCC cells. (A–C) Western Blot analysis of GRP78 expression in non-irradiated (0 Gy) and irradiated (6 Gy) HNSCC cells harvested with trypsin show no difference (A). Cells harvested with a cell scraper show an increase in GRP78 after irradiation (B). GRP78 expression was normalized to beta-actin and 0 Gy samples served as baseline (n = 4, t-test, *p < 0.05, **p < 0.01). (C) Representative Western Blot images of GRP78 expression in BHY and FaDu cells. Beta-actin was used as loading control. (D) Flow cytometry analysis of surface GRP78 in 0 Gy and 6 Gy-irradiated HNSCC cells confirmed increased GRP78 expression after irradiation. Samples were normalized to secondary antibody signal and 0 Gy (n = 4, t-test, *p < 0.05, **p < 0.01). (E, F) Example histograms of GRP78 fluorescence intensity from BHY cells harvested with either Trypsin (E), showing no difference in intensity, or a cell scraper (F), showing an increase of signal intensity in irradiated cells.