Las af v1

In situ hybridization chromogenic and immunofluorescence images were obtained with a DMCTR5000 microscope equipped with a DFC500 color camera (Leica. Wetzlar. Germany). Confocal microscopy was performed using a TCS-SP5 (Leica. Wetzlar. Germany) Laser Scanning System on Leica DMI8 microscopes. Up to 50 μm optical z-sections were obtained by taking 3.5 μm serial sections with LAS AF v1.8 software (Leica. Wetzlar. Germany). Tilescan mosaic images were reconstructed with Leica LAS AF software. All images were acquired using a 512 × 512 scan format with a 20 x objective. All coronal sections correspond to rostro-caudal coordinates between Bregma −0.82,–1.46 mm (Paxinos and Franklin, 2004 ). For the acquisition and quantifications of the fluorescence signal (Rodríguez-Tornos et al., 2016 (link); Briz et al., 2017 (link)), detectors were set to ensure equivalent threshold and signal-to-noise ratios between all samples. The maximum threshold signal was set by ensuring that no pixels were saturated. The threshold for background noise was determined using regions outside of the electroporated area (Rodríguez-Tornos et al., 2016 (link); Briz et al., 2017 (link)). This approach ensures linearity between samples.

Images were taken using a Leica DM4B (Leica) with a 2.5X objective and LAS AF v1.8 software (Leica; see Supplementary Materials for details).