Extended Data Fig. 10: Influence of NF157 on Shigella infection in CACO-2 cells and P2Y11 expression in corneal epithelial cells of guinea pigs.
a, Fluorescence microscopy analysis of influence of NF157 on the TSAR activation of Sf301 in non-polarized CACO-2 cells. F-actin is grey, Shigella bacteria are red, GFP-expressing bacteria are green and nuclei are stained with DAPI (blue). The bar represents 100 μm. b-d, Influence of NF157 on intracellular CFU (n = 6) (b), trans-epithelial electrical resistance (n = 3) (TEER, c) and epithelial integrity (d) of polarized CACO-2 cells infected with Shigella in the absence or presence of HD5. F-actin is red. Bacteria are green and nuclei are stained with DAPI (blue), and the bars represent 100 μm. Data are presented as mean±s.d. (b, c). Significance was determined by a two-tailed unpaired Student’ s t-test (b, c), and indicated as the p value. ns: no signification, **p < 0.01, ***p < 0.001 and ****p < 0.0001. e, Influence of NF157 on the Shigella T3SS activation (TSAR activity) on gut-on-chip with constant flow at the rate to 20 μL/h. The bars represent 50 μm. f. Immunofluorescence analysis of expression of P2Y11 in corneal epithelial cells of guinea pigs in the absence (upper panels) or presence (lower panels) of HD5 (Rhodamine-tagged). F-actin is red, P2Y11 is green, HD5 is grey and nuclei are stained with DAPI (blue), and the bars represent 100 μm. All results are representative of at least three independent experiments.
