Mouse anti blbp

The brains of E17.5 or P1 offspring mice were perfused and then post-fixed for 72 h with 4% PFA (pH7.4). The fixed mouse brains were cryoprotected in 30% sucrose solution and sectioned using a cryostat microtome at the thickness of 30 μm. The brain slices were washed with PBS and then incubated in 0.3% Triton X-100 for 30 min at room temperature. After blocking with 10% goat serum, the slices were incubated in primary antibodies at 4 °C overnight. The used primary antibodies were: mouse anti-BLBP (Abcam, 1:500), rat anti-Ctip2 (Abcam, 1:500), rabbit anti-Ki67 (Abcam, 1:500), rabbit anti-Pax6 (Covance, 1:1 000), mouse anti-Satb2 (Abcam, 1:1 000), rabbit anti-Sox2 (Millipore, 1:1 000), rabbit anti-Tbr2 (Abcam, 1:500), convalescent phase serum from the ZIKV patient (1:700). In the next day, the slices were incubated with Alexa Fluor 555/488/647 conjugated secondary antibodies (Invitrogen) for 2 h at room temperature. DAPI was obtained from Beyotime Biotechnology. Images were acquired using a Nikon A1 Plus confocal microscope with a 20× or 40× objective lens.