Figure 4
From: Discovery of rafoxanide as a novel agent for the treatment of non-small cell lung cancer
Rafoxanide promotes ERs and activates the UPR in A549 cells. (A) A549 cells were treated with 30 μM rafoxanide for 36 h. Cells were observed by electron microscopy. The ER (arrows) in cells is indicated. Scale bars, 500 nm. (B) A549 cells were treated with rafoxanide at different doses and cells were collected for western blot analysis at the indicated time points using antibodies against GRP78, pEIF2, and ATF6. β-actin was used as the loading control. The bar graph represents the quantitative analysis of western blots. (C,D) A549 cells were treated with rafoxanide (0, 7.5, 15, 30 μM) for 24, 36, 48 h. RNA was extracted from the treated and control cells. Both the unspliced and spliced forms of Xbp1 were analyzed by semi-quantitative RT-PCR. qRT-PCR was performed using primers specific for ATF4, GADD34, XBP1, EDEM2, ER57, and calnexin. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
