Fig 1: ITI-hUC-MSCs secrete C1INH to inhibit CD301+ macrophage polarization(A and B) The proportion of CD301+ macrophages and mean fluorescence intensity (MFI) of CD301 in PMA-treated THP-1 cells stimulated with LPS and IFN-? for 24 h and then treated with conditioned media (CM) from hUC-MSCs (n = 3) and ITI-hUC-MSCs (n = 3) for another 24 h was measured by flow cytometry.(C and D) Heatmap and volcano plot showing DEGs in hUC-MSCs (n = 3) and ITI-hUC-MSCs (n = 3). The red dots represent the upregulated transcripts, and the blue dots represent the downregulated transcripts (FDR (false discovery rate) < 0.05 and fold change >2).(E) Venn analysis of the top 200 genes by FC (fold change), top 200 genes by FDR and top 200 genes by expression abundance.(F) Heatmap showing the 43 genes filtered out through Venn analysis.(G) The mRNA levels of Serping1, Cxcl5, Il32, Il32a, Il32ß, Il32? and Mmp3 in hUC-MSCs (n = 4) and ITI-hUC-MSCs (n = 4) were measured by qRT?PCR.(H) The protein level of C1INH was examined by western blotting in hUC-MSCs (n = 3) and ITI-hUC-MSCs (n = 3).(I) The relative band intensities of C1INH were analyzed by ImageJ.(J and K) The proportion of CD301+ macrophages and MFI of CD301 in PMA-treated THP-1 cells stimulated with LPS and IFN-? for 24 h and treated with 100 ng/mL recombinant C1INH for another 24 h (n = 3) were measured by flow cytometry.(L) The mRNA level of Serping1 in hUC-MSCs (n = 5) transfected with three different small interfering RNA-seq of Serping1 (si-1, si-2, si-3) or negative control (si-NC) for 48 h was examined by qRT?PCR.(M) The protein level of C1INH in hUC-MSCs (n = 3) transfected with si-1, si-2, si-3 or si-NC for 48 h was examined by western blotting.(N) The relative band intensities of C1INH were analyzed by ImageJ.(O and P) The proportion of CD301+ macrophages and MFI of CD301 in PMA-treated THP-1 cells stimulated with LPS and IFN-? for 24 h and treated with conditioned media of ITI-hUC-MSCs transfected with si-NC (n = 3) and si-Serping1 (si-2; n = 3) for another 24 h was measured by flow cytometry. (A-B), (G), (I-L) and (N-P) Error bars indicate the mean ± SD. No significant difference (ns), *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.