Figure 3. T_reg directly enhance oligodendrocyte differentiation and myelination in vitro.

(a-d) Immunofluorescence analysis of (a) MBP⁺ cell numbers (t = 8.200, d.f. = 10, P < 0.0001) (b) percentage area (U = 0, P = 0.0022), (c) total Olig2⁺ cell numbers (t = 0.8294, d.f. = 10, P = 0.4263) and (d) representative images of mixed glial cultures analyzed (scale bar = 100 µm, green = Olig2, red = MBP), n = 6 wells.

(e,f) Immunofluorescence analysis of (e) Olig2⁺Ki67⁺ cell numbers (t = 1.299, d.f. = 10, P = 0.2230) with (f) representative images of mixed glial cultures (scale bar = 100 µm, green = Olig2, red = Ki67), n = 6 wells.

(g,h) Immunofluorescence analysis of (g) MBP⁺ cell numbers (t = 6.431, d.f. = 22, P < 0.0001) and (h) representative images of pure OPC cultures analyzed (scale bar = 50 µm, red = MBP, blue = DAPI), n = 12 fields of view.

(i,j) Immunofluorescence analysis of (i) MBP⁺CC1⁺cell numbers (n.d. = not detectable) and (j) representative images of DRG neuron-OPC co-cultures in the presence or absence of T_reg-conditioned medium, n = 20 fields of view. Image scale bars = 50 µm and 20 µm (enlarged image), red = MBP, green = CC1, blue = DAPI.

Data shown are representative of at least 12 (a,b,d), 3 (c,g-j) and 2 (e,f) independent experiments. Data presented with mean values indicated, error bars = SEM, Unpaired, two-tailed, Student’s t test (cell counts) and Mann-Whitney U tests (percentage area), ** p<0.01, *** p<0.001.