Fig. 5: HEK293T cells overexpressing mutant Cav3.2 channel show increased Ca²⁺ influx, hyperfunctional mTORC1 signaling, and impaired activation of a Reelin signaling component.

HEK293T cells were transfected with an empty vector (EMPTY-vector; n = 5) or with plasmids expressing either wild-type (WT-a1Ha; n = 5) or mutant (MUT-a1Ha; n = 5) Cav3.2 channels. A Intracellular Ca²⁺ measurements in transfected cells depolarized with 100 mM KCl and cultured in the absence (vehicle) or presence of 10 μM of the T-type Ca²⁺ channels blocker NNC 55-0396 (NNC). The bar graph shows the median value and interquartile range for each group. Cells overexpressing mutant Cav3.2 show significantly increased Ca²⁺ influx, which was rescued by treatment with NNC. Only the p-values obtained from the analyses using MUT-a1Ha are shown. B Representative immunoblots showing the expression levels of pRPS6, pSRC, SRC, and β-actin (used as loading control) in transfected cells cultured in the absence (−) or presence (+) of 10 μM of NNC. The bar graph shows the median levels and interquartile ranges of normalized pRPS6 and pSRC for each group; only the p-values obtained from the analyses using MUT-a1Ha are shown. Cells overexpressing mutant Cav3.2 show significantly increased pRPS6 levels and significantly decreased pSRC levels, which was rescued by treatment with NNC. *p < 0.05, **p ≤ 0.01, ****p ≤ 0.0001.