Figure 4

Inflammation induced CD14 in the embryonic intestinal stroma promotes inflammatory cytokine production and phagocytosis of monocyte derived cells. (A) CD14 mRNA expression normalized to Gapdh in LPS or vehicle treated adult and embryonic intestinal stromal cells, with the addition of control or CD14 siRNA knockdown in embryonic stromal cell cultures. n = 3 biological replicates. t-test. *p < 0.05, **p < 0.01, ****p < 0.0001. (B) IL1b and TNF ELISA using culture medium from LPS or vehicle treated adult and embryonic intestinal stromal cells, with the addition of control or CD14 siRNA knockdown in embryonic stromal cell cultures. n = 3 biological replicates. t-test. ****p < 0.0001 N.D.; not detected. (C) Representative fluorescent images of phagocytic cells identified with pHrodo red fluorescence in co-cultures of adult or embryonic stromal cells and MACS sorted GFP + monocytes. Scale bar = 100 µm. (D) Quantification of (C) showing the percentage of pHrodo red/GFP⁺ out of the total GFP⁺ cells in 2 HPF of 3 biological replicates. n = 3 biological replicates. t-test., ***p < 0.001, ****p < 0.0001. (E) Graphical abstract of the findings.