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. 2016 Dec 12;6:182. doi: 10.3389/fcimb.2016.00182

Figure 2.

Figure 2

ER stress is involved in M. bovis-mediated apoptosis. Raw 264.7 cells were infected with M. bovis at an MOI of 10, and then incubated for 0–48 h. (A) TEM analysis of Raw 264.7 cells after M. bovis infection for 6 h. The perinuclear rough ER regions on the images in the left panel are magnified on the right panels. Arrows indicate ER lamellae before and after infection. (B) XBP-1 mRNA splicing was determined by RT-PCR using specific primers that were used to amplify products of unspliced and spliced mRNA. (C) The results represent the ratio of spliced XBP-1 to intact (or unspliced) XBP-1 (XBP-1S/U ratio). (D) Total cell lysates were subjected to Western blot to identify phosphorylation of eIF2α. (E) Bands corresponding to each protein were quantified, and the intensities of each protein were normalized to the intensity of tubulin. Data are representative of at least three independent experiments, each performed in triplicate with similar results. The asterisks indicate significant differences compared with untreated cells (*P < 0.05, **P < 0.01, ***P < 0.001).