Fig. 6

MIF induces proliferation of BLEL-derived primary cells. (a) MIF increases BLEL primary cells proliferation in a time and dose-dependent manner. Cells from the same suspension were seeded at a density of 2 × 10³ cells per well in 96 well plates, let be adherent for 24 h and cultured with MIF (5 to 400 ng/ml), ISO-1 (100 μM) or without MIF nor ISO-1 at the indicated time point. Viable cells were counted using Cell Counting Kit-8 (CCK8). (b-d) MIF enhanced the expression of the proliferation marker Ki-67 in BLEL primary cells. Cells from the same suspension were seeded at a density of 2 × 10⁴ cells/well in a 24 wells plate, let be adherent for 24 h and cultured with MIF (200 ng/ml), ISO-1 (100 μM) or without MIF nor ISO-1 at the indicated time point: the percentage of Ki-67 positive cells counted using CellProfiler v2.2.0 (Cellprofiler™) were summarized for (b) BLELp1 cells (fibroblast) and (c) BLELp2 cells (myofibroblast); (d) representative images of the most significant time point were presented for Ki-67 immunostaining: BLELp1 (48 h) and BLELp2 (72 h). Representative results of three independent experiments were presented. Data were plotted as Mean ± SEM. * P-value < 0.05; ** P-value < 0.01; *** P-value < 0.001; no stars for P-value > 0.05. (d) Original magnification: 10×