Fig. 4

Cellular experiments showed LTBP1 could affect the function of GBM cells. a–b Western blotting was used to divide the primary GBM cell into high and low LTBP1 expression groups. β-actin was used as internal reference. *P<0.05; c The expression of LTBP1 were positively correlated with PHQ-9 and GAD-7 scores (n = 10). d Phase contrast pictures showed the morphology of spheres in both groups. Scale bar represented 200 mm. e The number of spheres in each well was counted on days 7, Data were presented as the mean ± SEM; Student’s t-test was chosen as the statistical method; n = 3, P = 0.0007 ***P < 0.001. f The diameter of spheres was measured to represent the volume. The diameters of spheres in both two groups increased gradually within 3, 5, 7 days, but the diameter of spheres in LTBP1 high expression group grew significantly faster. Data were presented as the mean ± SEM, one-way ANOVA was chosen as the statistical method. n = 3, *P < 0.05. g–h Cell cycle of two groups were detected by flow cytometry. A lower percentage of G1 phase, while higher percentage of S and G2-M phase were observed in high LTBP1 expression group than those in low LTBP1 expression group. **P < 0.01. ***P < 0.001. i–j Immunofluorescence showed more Ki-67 positive cells in high LTBP1 expression group than low LTBP1 expression group. n = 3, P = 0.0168. *, P < 0.05. k–l Wound healing assay showed faster migration capacity in LTBP1 high expression group than those in LTBP1 low expression group. *P < 0.05. m–n Transwell assay showed significantly better migration capacity in high LTBP1 group than those in Low LTBP1 group. n = 3, P = 0.0012