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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Investigation of de novo mutations in a schizophrenia case-parent trio by induced pluripotent stem cell-based in vitro disease modeling: convergence of schizophrenia- and autism-related cellular phenotypes

Fig. 4

Proliferation, scratch, and neurite outgrowth in the trio NPC lines. a For the proliferation assay, 35,000 NPCs were plated onto poly-ornithine/laminin coated plates and were further cultured for 4 days. Cells were harvested daily and cell number was measured by ATTUNE NXT flow cytometer. Non-viable cells were excluded by PI staining. Cell counts relative to seeded cell numbers (N/N0) are plotted per day. Values represent the means ± SE (N = 3 independent experiments, n = 3 technical replicates/experiments). * p < 0.05, **p < 0.01. b Representative images of scratch assay in NPCs. Three parallel scratches were made per dish by a sterile P5 pipet. Three photos were taken along every scratch, at 24 h. Manual analysis was performed using ImageJ. The rate of closure was defined by measuring the width of the scratches. c Analysis of the scratch closure relative to day 0. The diagram shows mean values ± SE (N = 2 independent experiments, n = 9 measurements/experiments). Scale bars = 100 μm. df Representative images of neurite outgrowth assay. NPCs were treated with DMSO or para-nitroblebbistatin (PNBS) (10 μM). The changes of neurite outgrowth were monitored and analyzed using the ImageXpress Micro XLS Widefield High-Content Analysis System. Diagrams show the mean neurite outgrowth ± SE of DMSO and PNBS treated NPCs visualized by Calcein after 4 h, normalized to 0 h (N = 3 independent experiments). Scale bars = 200 μm

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