Figure 8. HSP60 loss is associated with epithelial induction of WNT-related signals.

(a) IF images show epithelial RSPO1 (red) and HSP60 (yellow) expression at d₂ in jejunal sections from Hsp60^Δ/ΔIEC and Hsp60^flox/flox mice. E-Cadherin (grey) and αSMA (green) were used as epithelial and mesenchymal cell marker, respectively (DAPI stains nuclei in cyan, asterisks indicate Paneth cells). (b) IF images show epithelial WNT10A (red) and HSP60 (yellow) expression at d₂ in jejunal sections from Hsp60^Δ/ΔIEC and Hsp60^flox/flox mice. Lysozyme (green) and E-Cadherin (grey) were used as Paneth cell and epithelial cell marker, respectively (DAPI stains nuclei in cyan). WNT10A and Lysozyme co-stained (yellow, lower panel). (c) Quantification of WNT10A-positive cells in the jejunum of Hsp60^Δ/ΔIEC and Hsp60^flox/flox mice (N=6). Lines in the dot plot indicate mean numbers. Statistical analysis was performed using unpaired t-test. (d) Experimental scheme to induce Hsp60 loss in small intestinal organoids. qRT–PCR analysis of Lgr5 and Wnt10a mRNA expression in organoids following gene knockout. Data from organoid experiments derive from at least 3 independent experiments. Bars represent mean +s.e.m. One-way analysis of variance and appropriate post-hoc tests were used for statistical analysis. Asterisks in c, d indicate significant differences *P<0.05, **P<0.01, ***P<0.001; NS, not significant.