Fig. 4

Interplay between JAK/STAT and DLL4/Notch1 signaling. a, b, c Inhibition of JAK/STAT signaling pathway potentiates DLL4-mediated M2 apoptosis. Cells were polarized into M2 macrophages by IL-4 with or without the JAK/STAT inhibitor Ruxolitinib (RX) and with (+rhDLL4) or without (−rhDLL4) coated rhDLL4. a Expression of M2 markers at the membrane of M0 or M2 polarized macrophages with or without Ruxolitinib. Results from flow cytometry show geometric means of fluorescence expressed as relative expression (mean ± SD, n = 3). b Apoptosis of M2 polarized cells upon inhibition of JAK/STAT with Ruxonitilb (RX). After 24 h of polarization cells were stained with Annexin V/Propidium Iodide (IP). Results are representative dot plots from 3 independent experiments. c Histogram representing percentage of Annexin V positive cells (n = 3). d Flow cytometry of Notch1 surface expression on M1 and M2 differentiated for 48 h in the presence of immobilized DLL4. e Representative western blots for Notch1, N1ICD, HES1 and β actin in M1 and M2 Results are representative data from 3 independent experiments