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. 2021 Mar 30;11:7199. doi: 10.1038/s41598-021-86658-5

Figure 2.

Figure 2

ERp57 promotes hypoxic proliferation, is radioprotective and does not effect HIF signaling. (A) For colony formation assay, cells were seeded supplemented with doxycycline and incubated under normoxic or hypoxic conditions. After 24 h, cells were irradiated with 0, 1 and 3 Gy and further incubated for 10 d before formed colonies were counted. Survival fraction was presented in bar graphs and representative images of colonies formed in 6 well plates are shown. The seeded cell number is indicated for each well. (B) MTT assay 96 h after ERp57 KD (n = 10). (C) Protein analysis of HIF1α 72 h after ERp57 KD and 24 h of hypoxia by western blot. As a positive control for HIF1α expression, normoxic cells were treated with 2 mM of prolyl-4-hydroxylase inhibitor dimethyloxalylglycine (DMOG). (D) HIF reporter gene assay. After co-transfection with pGLHIF1.3 (firefly luciferase, 3 × HRE) and pGL4.74 (renilla luciferase), ERp57 KD was induced. After 24 h in normoxia, cells were incubated for 48 h in normoxia/ hypoxia before cell lysis (n = 4).