Fig. 4: Mutant Cav3.2-mediated hyperfunction of mTORC1 signaling acts alone to cause enhanced proliferation of F2688-1-derived NPCs and together with impaired Reelin signaling in the abnormal migration of these cells.

A Representative phase-contrast microscopy photographs of control- and F2688-1-derived NPCs. The bar graph shows the median cell body size (including soma size and all cell surface projections) of control-derived NPCs (n = 4) and F2688-1-derived NPCs (NPCs derived from 2 iPSC clones). There were no significant differences in median body size between the groups. B Line graph showing cell proliferation curves (live cell numbers at Day 0, 48 h, 72 h) of control-derived NPCs (n = 4) and F2688-1-derived (NPCs derived from 2 iPSC clones) cultured in the presence of either vehicle, 1 μM of NNC 55-0396 (NNC), or 100 nM of rapamycin; and in the presence of either mock-conditioned medium (mock) or Reelin-conditioned medium (Reelin). The graph shows the median value and interquartile range for each group. F2688-1 NPCs show significantly higher proliferation rates compared with control NPCs, which were completely rescued by treatment with either rapamycin or NNC. On the other hand, treatment with Reelin had no effect on the proliferation of F2688-1 NPCs. C Line graph showing the percentage relative wound density (RWD) over time in control-derived NPCs (n = 4) and in F2688-1-derived NPCs (NPCs derived from 2 iPSC clones) cultured in the presence of either vehicle, 1 μM of NNC, or 100 nM of rapamycin; and in the presence of either mock or Reelin. The graph shows the median value and interquartile range for each group. F2688-1 NPCs exhibit significantly higher migration rates compared with control NPCs, which was significantly attenuated by treatment with rapamycin, and completely rescued to levels equal to those of the untreated control NPCs in the exponential phase of the healing curve by treatment with either NNC or Reelin (analyzed time interval: 15 h–25 h). **p ≤ 0.01, ****p ≤ 0.0001.