AJCs are intact in the SNAP25 or Stx1A sgRNA electroporated region. (A) SNAP25 and Par3 stained in the electroporated region of the cerebral cortex. pX330 was electroporated as a control. mCherry staining shows the electroporated region. Lower panels (A′ and A′′) show magnified images of the boxes surrounded by dashed lines in the upper panels. Scale bars, 100 µm. (B) Magnified images of the neuronal layer in the boxes surrounded by dashed lines in the upper panel of A. SNAP25 staining was decreased in the electroporated region of the neuronal layer, indicating that Cas9 and sgRNA efficiently knocked out SNAP25. Scale bar, 100 µm. (C) Coimmunoprecipitation of Stx1A and Stx1B in the SNAP23 immunoprecipitates (IP) from the WT E13.5 brain lysate. (D) Stx1A and Par3 stained in the electroporated region of the cerebral cortex. pX330 was electroporated as a control. mCherry staining shows the electroporated region. Lower panels (D′ and D′′) show magnified images of the boxes surrounded by dashed lines in the upper panels. Scale bars, 100 µm. Ctrl, control.