Fig. 6: Characterization of neuronal phenotype. | Cell Death Discovery

Fig. 6: Characterization of neuronal phenotype.

From: A new advanced cellular model of functional cholinergic-like neurons developed by reprogramming the human SH-SY5Y neuroblastoma cell line

Fig. 6

A SH-SY5Y cells were plated and differentiated as described in the Materials and Methods section (3D DMAP2 Mix). Cells were fixed, permeabilized, and immunostained with either the anti-ChaT (green) or the anti-TH (orange) antibodies. A total of 10 z-stacks images for each condition were taken. Maximum projections of overlay are shown. The totality of fluorescence images was captured using Operetta CLS™ equipped with a 63× immersion objective. Scale bar: 50 µm. Quantification of (B) the ChaT/TH ratio in the cell body at 10 DIV (n = 528 cells) and 40 DIV (n = 951 cells). C The percentage of ChaT in the nucleus at 10 DIV (n = 433 cells) and 40 DIV (n = 743 cells) was performed using Harmony software, while quantification of (D) the number of ChaT and TH vesicles in neurites per 0.043 mm2 region at 10 DIV (ChaT n = 17 fields, TH n = 18 fields) and 40 DIV (ChaT n = 18 fields, TH = 17 fields) was performed using Fiji software. Statistical tests used to determine the significance of differences among the conditions were: Unpaired t-test, in panels B and C; Multiple t-tests, in panel D. Significance, was set as *p < 0.5, **p < 0.01, ***p < 0.001, ****p < 0.0001.

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