fig6

Figure 6. Increased CGN expression in glioblastoma contributes to decreased 3D growth. (A and B) Relative levels of miR-100 and miR-125b in CC, CC-CR, and LN-18 (glioblastoma) cell lines. Data represent mean ± SEM with ^** indicating P < 0.01 and ^**** indicating P < 0.0001. Significance was determined using Student’s t test; n = 3; (C) CGN levels were determined using western blots in CC, CC-CR, LN-18, and doxycyclin inducible LN-18 cell lines. Lane loading was monitored with blots against GAPDH; (D) Quantification of blots as in (C) showing fold changes in CGN in the indicated cell lines normalized to GAPDH. Data represent mean ± SEM with n = 3. Significance was determined using Student’s t test with ^**** indicating P < 0.0001; ns indicating not significant; (E) Increase in spheroid area was measured after transfer to collagen from 0-5 days. Surface area tracing was performed using ImageJ. Significance was determined using two-way ANOVA tests, n = 6. Data represent mean ± SEM with ^** indicating P < 0.01, ^*** indicating P < 0.001, ^**** indicating P < 0.0001; ns indicating not significant. CGN: Cingulin; CC: cystic colonies; SEM: scanning electron microscopy; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; ANOVA: analysis of variance.