Figure 2

PLD1 is involved in different signalling pathway triggered by BDNF in cultured cortical neurons.

(A) WT and Pld1^−/− cortical neurons in culture for 3 DIV were treated for the indicated time with 100 ng/mL of BDNF and lyzed. 35 μg of proteins was resolved by SDS-PAGE and probed with anti-ERK1/2, anti-phospho-ERK1/2 and anti-GAPDH antibodies. The phosphorylation level of ERK1/2 was corrected to ERK1/2 expression levels and subsequently normalized to GAPDH expression levels. The relative ERK1 and ERK2 is normalized to that of WT untreated cells obtained from three independent experiments. (B) WT and Pld1^−/− cortical in culture for 3 DIV were treated for 15 minutes with 100 ng/mL of BDNF and lyzed. 35 μg of proteins was resolved by SDS-PAGE and probed with anti-phospho-mTor (Ser-2481), anti-phospho-S6K (Thr-389), anti-phospho-CREB (Ser-133) and anti-GAPDH antibodies. The levels of phospho-proteins was normalized to GAPDH expression levels and normalized to that of WT untreated cells. For pS6K quantification, lower bands levels corresponding to P70S6K were analysed. Data were averaged from three independent experiments.