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. 2015 Dec 21;7(2):2080–2092. doi: 10.18632/oncotarget.6712

Figure 3. Activation of the ATF6 pathway in glioblastoma by ionizing radiation.

Figure 3

A. Luciferase assay for ATF6 transcriptional activity. D54 cells were co-transfected with p5xATF6-GL3 and siRNA targeting ATF6. The cells were irradiated with 6 Gy and assayed for luciferase activity 48h after IR. The relative activity normalized to control is shown. B. qRT-PCR analysis of HERPUD1 and GRP78 expression in D54 after knockdown of ATF6. Cells were treated with ATF6 siRNA for 48h prior to irradiation with 6 Gy, and harvested 48h later. C. Western blot analysis of GRP78 in whole-cell lysates from D54 cells harvested at the indicated time-points after irradiation with 6 Gy. D. Western blot analysis of GRP78 in whole-cell lysates from D54 cells harvested 48h after irradiation with 3 and 6 Gy. E. Western blot analysis of GRP78 in whole-cell lysates from D54 cells treated with ATF6 siRNA and 6 Gy IR. Cells were harvested 48h after irradiation. Shown beneath the blots is the relative expression of the indicated proteins when compared to the respective untreated control. The values represent the normalized densitometric output for each band divided by the corresponding loading control. In all graphs, data shown are the Means ±SD (n = 3). *P < 0.05, ***P < 0.001, ****P < 0.0001.