Fig. 3

EndoMT-derived cells stimulate macrophage infiltration and M2-polarization. a Set-ups of transendothelial migration assays using mouse (upper panel) or human (bottom panel) cells. b Transendothelial migration activities of non-polarized, M1-polarized, and M2-polarized RAW264.7 cells upon co-culture with Panc 02 cells alone or Panc 02 cells plus 3B-11 cells or OPN-treated 3B-11 cells. ^αP < 0.01 when compared with “Panc 02” group. ^βP < 0.01 when compared with “M0” group. c Transendothelial migration activities of non-polarized, M1-polarized, and M2-polarized differentiated THP-1 cells upon co-culture with PANC-1 cells alone or PANC-1 cells plus HUVECs or OPN-treated HUVECs. ^αP < 0.05 and ^βP < 0.01 when compared with “PANC-1” group. ^γP < 0.05 and ^δP < 0.01 when compared with “M0” group. d mRNA levels of IL-1β, TNF-α, CD163, CD204, IL-10, and TGF-β in the THP-1-derived macrophages treated 24 h with control medium (Ctrl), Endo CM, or EndoMT CM. *P < 0.05 and ^#P < 0.01 when compared with “Ctrl” group. e Secreted levels of IL-1β, IL-10, and TGF-β from the THP-1-derived macrophages treated 24 h with Ctrl, Endo CM, or EndoMT CM. THP-1-derived macrophages were pre-incubated with 1% FBS-containing RPMI 1640 medium for 16 h. The medium was then replaced with Ctrl, Endo CM, or EndoMT CM for further 24 h. The treated macrophages were further incubated with 5 ml of fresh 1% FBS-containing medium for 24 h. The media were finally collected for ELISAs. ^#P < 0.01 when compared with “Ctrl” group. f Cell-surface levels of CD163 and CD204 in the THP-1-derived macrophages treated 24 h with Ctrl, Endo CM, or EndoMT CM. g mRNA expression status of Arg1 and iNOS in the THP-1-derived macrophages treated 24 h with Ctrl, Endo CM, or EndoMT CM. h mRNA levels of IL-1β, TNF-α, iNOS, CD163, CD204, IL-10, TGF-β, and Arg1 in the THP-1-derived macrophages treated 24 h with LPS plus IFN-γ (M1-inducer) in control medium (Ctrl) or EndoMT CM. *P < 0.05 and ^#P < 0.01 when compared with “LPS + IFN-γ + Ctrl” group. i mRNA levels of IL-1β, TNF-α, iNOS, CD163, CD204, IL-10, TGF-β, and Arg1 in the THP-1-derived macrophages pretreated with LPS plus IFN-γ for 6 h and treated with control medium (Ctrl) or EndoMT CM containing fresh LPS and IFN-γ for further 24 h. *P < 0.05 and ^#P < 0.01 when compared with “LPS + IFN-γ ➔ Ctrl” group