A, Images of Western blot analysis of c-Jun and MAP-LC3β expression during Chlamydial infection. C57epi cells infected with C. muridarum (MoPn) induced the upregulation of key proteins in the UPR pathways, including c-Jun and MAP-LC3β and inhibition of IRE1 RNase activity result in their downregulation. Ten μg total protein from C57epi cells uninfected/infected with C. muridarum for 48 hours and treated/nontreated with inhibitor of IRE1α RNase activity were prepared for Western blot analysis. Blot was probed with primary antibody against c-Jun, Glut1, hexokinase II, and MAP-LC3β. Lane 1: protein molecular weight marker. Lane 2: sample from noninfected C57epi cells at 48 hours. Lane 3: sample from C57epi cells infected with MoPn at MOI 5 for 48 hours; Lane 4: Sample from C57epi cells infected with MoPn at MOI 5 in the presence of inhibitor of IRE1α RNase activity for 48 hours. B, Western blot of MAP-LC3β. Ten μg total protein from cells infected with C. muridarum (MoPn) for 24, 48, and 72 hours were prepared for Western blot analysis. Blot was probed with primary antibody against MAP-LC3β. Lane 1: protein molecular weight marker. Lanes 2–4: samples from noninfected C57epi cells at 24, 48, and 72 hours. Lanes 5–7: samples from C57epi cells infected with C. muridarum (MOI 1) at 24, 48, and 72 hours. Lanes 8–10: samples from C57epi cells infected with C. muridarum (MOI 5) at 24, 48, and 72 hours. Abbreviations: C57epi cells, mouse oviduct epithelial cells; Glut1, glucose transporter-1; IRE1α, inositol-requiring enzyme-1α; MAP-LC3β, microtubule-associated protein 1 light chain 3; MOI, multiplicity of infection; MoPn, mouse pneumonitis; NI, noninfected; RNase, ribonuclease.