Fig. 7: Effect of the chimeric protein XIP-NLS on [Ca²⁺]_n and neuronal differentiation in primary cortical neurons.

a Western blot of NCX1, Lamin B1/B2, and karyopherin in inner and outer NE membranes of cortical neurons at 7 DIV. All the experiments were repeated at least three times on different preparations. b Nuclear lysates from neurons trasfected with XIP-NLS (48 h) and immunoprecipitated with anti-EYFP (top) or anti-NCX1 (bottom). The presence of NCX1 (top) or EYFP (bottom) was analyzed by immunoblotting performed with specific antibodies. The input consists on nuclear lysates from untreated neurons at 7 DIV. c Representative traces of Fura-dextran-detected nucleoplasmic Ca²⁺ after ATP (100 µM) perfusion in cortical neurons previously transfected with mock or XIP-NLS. Bar graph depicts quantification of ATP-induced nucleoplasmic Ca²⁺ increase. *p < 0.05 vs. mock. mock (n = 15 cells); XIP-NLS (n = 10 cells). d, e Representative Western blots and quantifications of GAP-43, MAP-2, PTEN protein expression, and Akt phosphorylation in cortical neurons (7DIV) previously transfected with mock or XIP-NLS. All the experiments were repeated at least three times on different preparations; *p < 0.05 vs. mock