Fig. 7: IEC overexpression of SIRT6 elevates the Tyr641 phosphorylation of STAT6 and enhances anti-helminth responses. | Nature Communications

Fig. 7: IEC overexpression of SIRT6 elevates the Tyr641 phosphorylation of STAT6 and enhances anti-helminth responses.

From: Sirtuin 6 maintains epithelial STAT6 activity to support intestinal tuft cell development and type 2 immunity

Fig. 7

3-month-old male WT and IEC-Tg mice were infected with H. poly and subjected to the following analyses on day 14 post-infection. a, b Western blot (a) and quantification (b) analyses of SIRT6 and P-STAT6 (Y641) in the jejunal IECs. n = 4 mice/group (SIRT6) and 5 mice/group (P-STAT6); p = 0.0057 (SIRT6, P-STAT6). c Tuft and goblet cells were examined by DCLK1 immunostaining and Alcian blue staining, respectively in the jejunum (200X). d Quantification of tuft and goblet cells shown in (c). n = 4 mice/group; 20 crypt-villus units counted for each mouse; p = 0.0118 (tuft cell) and 0.0338 (goblet cell). e qPCR analysis of tuft and goblet cell markers expression in the jejunal IECs. Dclk1, n = 8 (WT) and 7 (IEC-Tg) mice; Trpm5, n = 6 (WT) and 7 (IEC-Tg) mice; Retnlb, n = 7 mice/group; p = 0.0489 (Dclk1). f qPCR analysis of type 2 immune cytokine genes expression in the jejunum. Il4, n = 7 mice/group; Il5, Il13, n = 8 (WT) and 7 (IEC-Tg) mice; Il9, n = 6 mice/group; p = 0.028 (Il5). g Jejunal IL13 levels were measured by ELISA. n = 6 mice/group. h, i Analysis of parasite burden by quantification of adult worms in intestinal lumen (h) and eggs in feces (i). n = 8 mice/group; p = 0.0227 in (h). j Tuft and goblet cells were labeled by anti-DCLK1 and anti-MUC2, respectively, in frozen sections of rIL13-treated (25 ng/ml, 48 h) organoids (200X). k Quantification of tuft and goblet cells shown in (j). n = 14 (tuft cell) and 12 (goblet cell) organoids/group; p = 0.0029 (tuft cell) and 0.0451 (goblet cell). l qPCR analysis of the expression of Dclk1, Trpm5 and Retnlb in organoids treated with rIL13. n = 4 biological replicates/group; p = 0.0145 (Dclk1), 0.0057 (Trpm5), 0.0237 (Retnlb). m Immunostaining for P-STAT6 (Y641) in the jejunum. n, o Western blot (n) and quantification (o) analyses of P-STAT6 (Y641) in rIL13 treated organoids. n = 3 biological replicates/group. Data are presented as mean ± SEM. All p values were generated by two-tailed unpaired t test. *p < 0.05. Scale bars, 100 µm in (c, m); 50 µm in (j). Source data are provided as a Source Data file.

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