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. 2017 Jul 6;7:4787. doi: 10.1038/s41598-017-05305-0

Figure 3.

Figure 3

Overexpression of Znf179 in primary glioblastoma multiforme (GBM) cells promotes neuronal differentiation accompanied by the induction of the glial marker, glial fibrillary acidic protein (GFAP). Primary GBM-derived cells stably expressing Flag-Znf179 (A) or GFP-Znf179 (B) were differentiated in DMEM/F12 medium containing 1% FBS as described in “Materials and Methods”. Average numbers of glioma tumor spheres and sphere sizes of GBM cells were determined in at least three independent experiments. Scale bar: 20 μm. *p < 0.05; **p < 0.01; ***p < 0.001 (groups were compared by t-test, two-tailed p values). (C) Primary GBM-derived cells stably expressing Flag-Znf79 or GFP-Znf179 were differentiated and immunostained with an anti-GFAP antibody. The GFAP(+) differentiated GBM cells with significant long neurite branching were shown in red. Quantification of the percentage of GFAP(+) cells is shown in the histogram. ***p < 0.001 (groups were compared by a t-test, two-tailed p values). Scale bar: 50 μm. (D) Total lysates of these GBM cells were analyzed by Western blotting using anti-Znf179, anti-CD133, anti-nestin, anti-GFAP, and anti-β-actin antibodies. Uncropped images are in Supplementary Figure S3.