Figure 3

Assessment of the gliogenic-suppression effect of ruxolitinib in differentiating neurospheres ex vivo. (A) A schematic diagram depicting the study outline of ex vivo differentiation of neurospheres for ruxolitinib treatment. (B) Immunoblotting analysis for total Jak2, phosphorylated Jak2 (n = 2 per group), Tuj1 and Gfap (n = 2 per group) in vehicle control (G1) and all treated groups (G2-G7). The corresponding pixelation analysis of immunoblotting bands is shown below to the blots (n = 2 per group; Welsh's t-test). (C) Immunocytochemistry staining of Tuj1 and Gfap in differentiating neurospheres in control and 10 μM ruxolitinib treated groups (n = 2 per group). The higher magnification of insets in the control group is shown at the bottom of the micrographs. The colour of the insets corresponds to the DAPI (blue), Tuj1 (green) or Gfap (red). Scale bars represent 200 μm in all micrographs whereas 20 μm in the insets. All values represent mean ± SEM; * denotes adjusted p < 0.05; ** denotes adjusted p < 0.01. See Fig. S4 for original immunoblots (Supplementary information).