Figure 4
Hspa5 is absent in the dysplasia of the K19-wnt1/C2mE mice but is upregulated in the dysplasia of H. felis-infected mice and colocalizes with alcianophilia in spasmolytic polypeptide expressing metaplasia (SPEM). (a) Increased Hspa5 expression (left) within SPEM (area enclosed in circle) and dysplasia (area enclosed in square) induced by 78-week infection of C57Bl/6 mice with H. felis. Co-localization of Hspa5 with alcianophilia in SPEM is depicted by PAS/AB staining of serial section. (a, inset) Increased Hspa5 expression (left panel) on the apical surface of epithelial cells in dysplasia and in the inflammatory infiltrates (arrow). PAS/AB staining of serial section showing lack of Hspa5 co-localization with alcianophilia in the dysplastic region (right). (b) Minimal Hspa5 is expressed in the mucus metaplasia and dysplasia of the K19-wnt1/C2ME mice. Similar results were obtained for the K19-C2mE mice (data not shown). PAS/AB staining of the corresponding section shows lack of alcianophilia in the metaplastic and dysplastic region. (b, inset) Higher magnification of the same section (area enclosed in square) depicting low level of Hspa5 in the dysplastic and metaplastic cells and alcianophilic goblet cells. (c) Quantitative real-time PCR (qPCR) measurement of Xbp1-u, Xbp1-s, Hspa5 and Chop mRNAs in the H. felis-infected C57Bl/6 mice, uninfected K19-Wnt1/C2mE and K19/C2mE mice. RNA levels are expressed as fold expression normalized to β-actin, relative to uninfected wild-type C57Bl/6 mice. *Hspa5 level was significantly increased in the H. felis-infected mice compared with the K19-wnt1/C2ME mice (P=0.04). #Chop was significantly higher in the H. felis-infected mice compared with the K19-Wnt1/C2ME mice (P=0.02) or K19-C2ME mice (P=0.01). (d) Average qPCR measurement of Xbp1-u, Xbp1-s, Hspa5 and Chop mRNAs in NIH/3T3 cells treated with 1.5 μmol/l TG for 4 h relative to untreated cells. All data are represented as mean±s.e. from triplicate experiments.
