Journal: Immunology

Article Title: Acute infection of mice with Clostridium difficile leads to eIF2α phosphorylation and pro-survival signalling as part of the mucosal inflammatory response.

doi: 10.1111/imm.12122

Figure Lengend Snippet: Figure 6. Assessment of the unfolded protein response in Clostridium difficile-infected (CDI) mice. Protein lysates from the caeca and colons of untreated and C. difficile-infected mice were used to evaluate the phosphorylation of eIF2a (n = 6 pairs) (a), and the expression levels of BiP (n = 6 pairs) (d), P58IPK (n = 6 pairs) (e) and calreticulin (n = 6 pairs) (f). In each case, the panel on the left shows the image of the immunoblot for the evaluated molecule for three pairs of caeca and colons and the bar graph on the right depicts the mean SEM of the response for all the six evaluated pairs. The expression levels of Gadd34 and Wars were determined by quantitative RT-PCR (n = 12 pairs). Each bar represents the mean SEM of the relative expression of the depicted gene (b). Conventional RT-PCR was used to determine the lack of Xbp1 splicing (n = 12 pairs, of which 6 are shown) (c). A P-value ≤0.05 indicates a significant difference between the untreated and CDI samples and is marked with a ⋆.

Article Snippet: Immunoblotting antibodies against b-actin (clone 13E5), calreticulin, phospho-eIF2a (clone 119A11), eIF2a (clone L57A5), GAPDH (clone 14C10), P58IPK (clone C56E7), phospho-AKT (clone D9E), AKT (clone C67E7), phospho-STAT3 (clone D3A7) and STAT3 (clone 79D7) were obtained from Cell Signaling Technology (Danvers, MA).

Techniques: Infection, Phospho-proteomics, Expressing, Western Blot, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction