Fig. 5.

Ca²⁺ imaging in endoplasmic reticulum (ER) of control and TSC astrocytes. a. Control and TSC astrocytes, previously loaded with Fura-2/AM, were stimulated with TBHQ (50 s) and Ca²⁺ (300 s) in KRB solution. Representative curves of cytosolic Ca²⁺ responses upon the applied stimulus of three independent experiments. The blue curve showed the Ca²⁺ response of control astrocytes, while the dark orange curve showed the Ca²⁺ response of TSC astrocytes (n = 160 control cells and n = 136 TSC cells, from 14 independent coverslips). b. Bar plot visualizing significant reduction of Ca²⁺ concentration in the cytoplasm of TSC astrocytes compared to control astrocytes after blocking the SERCA protein with TBHQ stimuli (between 30 and 300 s). c. Bar plot visualizing basal cytosolic Ca²⁺ concentration in TSC astrocytes after TBHQ stimuli compared to controls. d. Bar plot visualizing significant reduction in the absolute concentration of Ca²⁺ in the ER of TSC astrocytes compared to control astrocytes after Ca²⁺ administration (at 300 s). Data are represented as mean ± SEM; p value: *p value ≤ 0.05; **p value ≤ 0.01; ***p value ≤ 0.001; ****p value ≤ 0.0001. Mann–Whitney U tests were used to evaluate significance