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. 2022 Sep 3;13:5192. doi: 10.1038/s41467-022-32846-4

Fig. 1. IEC-specific deletion of Sirt6 results in defective tuft cell expansion and intestinal type 2 immune responses.

Fig. 1

2–3-month-old male naive and/or H. poly-infected LoxP and IEC-KO mice were subjected to the following assays on day 14 post-infection. a, b Western blot (a) and qPCR (b) analyses of SIRT6 in the jejunal and colonic IECs of naive mice. n = 5 mice/group in (b); p = 0.0028 (jejunum) and 0.0014 (colon). c, d qPCR analysis of the expression of IEC markers in the jejunal (c) and colonic (d) IECs of naive mice. Dclk1, Muc2, Slc5a1, Chga, Lyz1, n = 5 mice/group; Lgr5, n = 4 mice/group; p = 0.0096 (Dclk1) in (c). e Jejunal tuft cells were examined by immunostaining with anti-DCLK1 antibody (200X). f Jejunal goblet cells were examined by Alcian blue staining (200X). g Quantification of tuft (DCLK1 positive) cells shown in (e). n = 4, 4, 5, 4 mice/group, respectively; 15 crypt-villus units counted for each mouse; from left to right, *p = 0.0188, <0.0001, 0.0093; #p < 0.0001 for both. h Quantification of goblet (Alcian blue positive) cells shown in (f). n = 4, 4, 5, 4 mice/group, respectively; 15 crypt-villus units counted for each mouse; from left to right, *p = 0.0005; #p = 0.0001, 0.0021. i, j Analysis of parasite burden by quantification of adult worms in intestinal lumen (i) and eggs in feces (j). n = 5 mice/group; p = 0.0243 in (i) and 0.0029 in (j). k qPCR analysis of tuft and goblet cell markers expression in the jejunal IECs. n = 4, 4, 5, 5 mice/group, respectively; from left to right, Dclk1 (*p = 0.0497, 0.0004; #p = 0.0005, 0.0002); Il25 (*p = 0.0474, <0.0001; #p < 0.0001, 0.0006); Pou2f3 (*p = 0.032, 0.0007; #p = 0.0006, 0.0004); Muc2 (*p = 0.0296, 0.0466; #p = 0.0214); Retnlb (*p = 0.0016; #p = 0.0019, 0.0018). l Representative FACS plots of H. poly-infected mice analyzed for IL17RB+CD90+ ILC2s gated from Lin−CD45+CD3−CD4− cells in the intestinal lamina propria. n = 4 mice/group. m qPCR analysis of type 2 cytokines expression in the jejunum of mice infected with H. poly. n = 10, 6, 6, 11 mice/group for Il4, Il5, Il9, Il13, respectively; p = 0.0484 (Il4) and 0.0269 (Il13). n Jejunal IL13 levels were measured by ELISA. n = 7 mice/group; *p = 0.0114; #p = 0.0072. Data are presented as mean ± SEM. All p values were generated by two-tailed unpaired t test. *p < 0.05. In (g, h, k, n), *p < 0.05 vs LoxP naive, #p < 0.05 vs LoxP H. poly. Scale bars, 100 µm. Source data are provided as a Source Data file.