Fig. 3 | Nature Communications

Fig. 3

From: Analysis of chromatin accessibility uncovers TEAD1 as a regulator of migration in human glioblastoma

Fig. 3

CRISPR-Cas9 ablation of TEAD1/4 inhibits migration in primary GBM cells. a Western immunoblot confirms population knockout of TEAD1 and TEAD4 after CRISPR-Cas9-mediated gene ablation. b Cell growth analysis reveals significantly decreased proliferation in TEAD1KO cells at 48–72 h, compared to sham (n = 3; 48 h: **p = 0.008; 72 h: *p = 0.01. Bars represent mean ± SEM). c Neurosphere (NS) assays show no difference in sphere number (day 6; n = 3 wells, multiple NS per well. Bars represent mean ± SEM). d Neurosphere (NS) assays show decreased sphere size in TEAD1 knockout, compared to sham. (day6; n = 3 wells, multiple NS per well; TEAD1KO: **p = 0.002. Dots represent individual NS and lines delineate mean). e Transwell invasion assays show decreased percent cell invasion in TEAD1KO and TEAD4KO cells, compared to sham (24 h; n = 3 wells; TEAD1KO: **p = 0.002; TEAD4KO: **p = 0.001. Bars represent mean ± SEM). On right, representative images of transwell invasion chamber membranes are shown. f, g Spheroid migration assays show decreased area of confluent cell migration (dispersion) at 36 h in TEAD1KO and TEAD4KO cells, compared to sham (f, PDL substrate), with partial rescue of migratory deficits in TEAD1KO cells after TEAD1 overexpression (OE) (g, laminin + PDL substrate) (n = 3 wells, with multiple NS per well; TEAD1KO: ***p = 0.0001; TEAD4KO: ***p = 0.00007; TEAD1OE 1: ***p = 0.00058; TEAD1OE 2 (1/10 dilution of TEAD1OE 1): ***p = 0.00015. Bars represent mean ± SEM). On right, migration area marked by red dash line in representative spheroids is shown. All experiments in ag are performed on G-13063 cells. Scale bars = 75 μM. See also Supplementary Fig. 5

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