Ttyh1 deficiency slows glioma-cell invasion. a, b, Time series of 3D images of S24 shControl GBMSCs (a; z = 36 μm) and S24 shTtyh1 GBMSCs (b; z = 69 μm) reveal the impaired invasion capacity induced by Ttyh1 knockdown. Left, Arrows indicate the subsequent invasion route; arrowheads label the cell bodies of the invading GBMSCs (green). Red, blood vessels. c, Measurements of the invasion speed of single S24 shControl versus shTtyh1 GBMSCs in vivo measured by repetitive MPLSM over 24 h (128–150 cells from n = 3 mice per group; day 21 ± 1; Mann–Whitney test). d, Measurements of the invasion speed of S24 shControl versus shTtyh1 GBMSCs in vivo subcategorized regarding their number of TMs (13–126 cells from n = 3 mice per group; day 21 ± 1; ANOVA on ranks). e, 3D confocal microscopy image of an immunocytochemical staining of integrin-α5 (ITGA5; green) and Ttyh1 (red) in S24 cells (arrow, ITGA5 enrichment at the growth cone-like tip; arrowhead, colocalization of ITGA5 and Ttyh1 at the tip of the TM). a–d, Data obtained by in vivo MPLSM. *p < 0.05; ***p < 0.001.