Fig 2: Decreased infiltration of particular IC subsets in the colons of the b1/2-ARs KO chimera. Significantly lower levels of CD4+ cells (A), CD11b+ macrophages (B) and CD94+ NK cells (C) were observed in the colons of b1/2-ARs KO chimeras compared with C57 control chimeras. Quantified data were presented in the (D). Blue indicates non-specific DAPI staining, while red shows CD4-, CD11b-, and CD94- positive cells, respectively. Images were obtained at 400X magnification. All bars indicate 20 µm. Unpaired two tailed t-test showed ***P < 0.001 and ****P < 0.0001.

Fig 3: Zymosan-treated human serum-induced TNFa production in neutrophil-like HL60 cells.HL60 cells were induced to differentiate into neutrophil-like cells by treatment with 2.5 µM ATRA for 0–5 days. (A) Flow cytometric analysis of the cell surface expression of CR3, C3aR, or C5aR1 in day 0, 1, 3, and 5 ATRA-treated HL60 cells. (B) Day 0–5 ATRA-treated HL60 cells were either untreated (NT) or incubated with 10% human serum or zymosan-treated human serum for 4 h, and the whole cell lysates (Cell) and culture supernatants (Sup.) were then collected. The protein samples were separated in SDS-PAGE, and TNFa and ß-actin were detected by immunoblot analysis. RPMI, Human serum, and ZHS in diagonal letters indicate samples of RPMI-1640 medium, RPMI-1640 containing 10% zymosan-treated human serum, or human serum mixed with 1 × SDS-PAGE sample loading buffer, respectively. (C, D) Day 5 ATRA-treated HL60 cells were either untreated (NT) or incubated with 10% human serum or zymosan-treated human serum for indicated time points, and the whole cell lysates (Cell) and culture supernatants (Sup.) were then collected. (E) Day 5 ATRA-treated HL60 cells and PMNs were either untreated (NT) or incubated with 10% human serum (HS) or zymosan-treated human serum (ZHS) for 4 h, and the whole cell lysates (Cell) and culture supernatants (Sup.) were then collected. The protein samples were separated in SDS-PAGE, and TNFa and ß-actin were detected by immunoblot analysis. The blot is a representative of 3 independent experiments.