Fig. 3S.Neuroblast-like characteristics of CN. (a, b) Representative images of MRI (Left), H&E (Middle), and Synaptophysin or NeuN (Right) of two patients among three patients. A T2 weighted brain MRI shows an inhomogeneous mass lesion in the left lateral ventricle of Patient 1 (a) and both lateral ventricle of Patient 2 (b). Patient 2 has much bigger size of tumor mass when compared to Patient 1. Red dotted line indicates CN tumor mass. H&E of the Patient 1 (a) and 2 (b) shows sheet of monotonous rounded cells with rounded nuclei. Synaptophysin (a) and NeuN (b) is robustly positive in tumor cells of the patients. (c, d) CN tissue with immunofluorescence staining of PSA-NCAM, BMP2, and Tuj1 (c) and NeuN and MAP2 (d). Each first panel indicates co-localization. (e) Immunofluorescence staining of CN-derived cells grown in ITSFn media shows localization of PSA-NCAM, BMP2 and Tuj1 representing neuroblast character. (f) CN-derived cells have round shape without mature neuronal structures. (g) Up: RT-PCR of CN tissue and CN-differentiated cells shows expression of BMP2, BMP4, BMPR1B, NCAM, Tuj1, and GAPDH. Bottom: Densitometry measurement of each genes relative to GAPDH in tissue and cell. F: 10.77, DFn: 4, p<0.001; Row factor has significant effects. (h) A few cells in CN tissue are Nestin+/BLBP+ representing RGC. (i) CN tissue has a mixed population of BLBP+ cells, BLBP+/PSA-NCAM+, and PSA-NCAM+ cells. (j) Left: RT-PCR of CN tissue and CN-differentiated cells detected expression of BLBP, Nestin, GFAP, and GAPDH. Right: Densitometry measurement of each genes relative to GAPDH. F:2.06, DFn:1 p=0.1461 no effect. Original full-length images of g and j are presented in Supplementary Scale bar 6 cm (a), 50 µm (c) 20 µm (e) 100 µm (h, i) 20 µm. All data presented as mean +/- S.D. Two way ANOVA followed by Bonferroni correction and t-test analysis was done. A p value of <0.01 (**), 0.001 (***), and ns: not significant was used to denote statistical significance. Three biological samples (N=3) were used for the experiments.
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