After washing in PBS, sections or fixed cells (with PFA 4%, 10 min at room temperature) were treated with PBS - 0.01% Triton X100 - 10% serum for 30 min. They were then incubated overnight at 4°C with the following primary antibodies diluted in the blocking buffer: rabbit anti-CaMKIIβ (1/300; Abcam, ab34703), mouse anti-CaMKIIδ (1/50, Santa Cruz, sc-100362), mouse anti-CaMKIIγ (1/50, Santa Cruz, sc-517278), rabbit anti-Cux1 (1/50; Santa Cruz, sc-13024), chicken anti-GFP (1/1000; Abcam, ab13970), mouse anti-Ki67 (1/50; BD Pharmingen™, 550609), mouse anti-NeuN (1/500; Millipore, MAB377), rabbit anti-pHH3 (1/200; Upstate, 06-570), rabbit anti-Tbr2 (1/500; Abcam, ab23345), mouse anti-Tuj1 (1/200; Covance, MMS-435P). Sections were then incubated with appropriate fluorescent secondary antibodies. For Cux1 and NeuN immunostainings, a step of antigen retrieval with citrate was performed before the step of blocking (sodium citrate pH=6 for 15 min at 90°C).
F-actin filaments were visualized using rhodamine-labelled phalloidin (Sigma). After fixation, sections or cells were permeabilized 10 min with 0.1% Triton X100 - PBS, incubated with rhodamine-labelled phalloidin diluted in PBS (0.2 μg/ml) for 40 min and washed several times in PBS.
F-actin filaments were visualized using rhodamine-labelled phalloidin (Sigma). After fixation, sections or cells were permeabilized 10 min with 0.1% Triton X100 - PBS, incubated with rhodamine-labelled phalloidin diluted in PBS (0.2 μg/ml) for 40 min and washed several times in PBS.