The influence on Muc2 positive cells and Gli1 positive cells after the knockout of Smad4 or Alk3 in Gli1 positive cell and the effect of IL1 or IL17 on colonic organoids. The influence on Muc2 positive cells and Gli1 positive cells after the knockout of Smad4 or Alk3 in Gli1 positive cell and the effect of IL1 or IL17 on colonic organoids

To investigate the influecne of BMP signaling on Gli1+ cells, RNA-seq were performed after Smad4 or Alk3 knockout. Next, RNA-seq of Muc2-mCherry cells were performed to reveal the regulation of Gli1+ cell on intestinal epithelium. Finally, RNA-seq of colonic organoids were performed to reveal the regulation of IL-1 or IL-17 on epithelium. To investigate the influecne of BMP signaling on Gli1+ cells, RNA-seq were performed after Smad4 or Alk3 knockout. Next, RNA-seq of Muc2-mCherry cells were performed to reveal the regulation of Gli1+ cell on intestinal epithelium. Finally, RNA-seq of colonic organoids were performed to reveal the regulation of IL-1 or IL-17 on epithelium. Overall design: One monthe after tamoxifen administration, FACS was performed to sort Muc2-mCherry or Gli1-tdTomato cells and then performed RNA sequencing. Alao, RNA-seq were performed in colonic organoids at 12 or 72 hours after IL-1 or IL-17 treatments.

为探究骨形态发生蛋白（BMP）信号通路对Gli1+细胞的影响，分别在Smad4或Alk3基因敲除后开展RNA测序（RNA-seq）；随后，对Muc2-mCherry阳性细胞进行RNA测序，以揭示Gli1+细胞对肠上皮的调控作用；最后，对结肠类器官实施RNA测序，以解析白细胞介素1（IL-1）及白细胞介素17（IL-17）对上皮细胞的调控效应。整体实验设计：在他莫昔芬给药1个月后，通过荧光激活细胞分选（Fluorescence-Activated Cell Sorting, FACS）分选出Muc2-mCherry或Gli1-tdTomato阳性细胞，随后进行RNA测序；此外，在IL-1或IL-17处理结肠类器官12小时或72小时后，同样开展RNA测序。