Chicken anti mbp

To detect RABV in the tissues, a polyclonal rabbit serum against recombinant RABV P protein (P160-5; 1:3000 in PTwH [0.2% Tween 20 in PBS with 10 µg/mL heparin]) was used [45 (link)]. The following first antibodies and dyes were obtained from the respective suppliers: chicken anti-NEFM (Thermo Fisher, USA; #PA1-16758, RRID:AB_2282551; 1:1000 in PTwH), guinea pig anti-NEFM (Synaptic Systems, Germany; #171204, RRID:AB_2619872; 1:400 in PTwH), chicken anti-MBP (Thermo Fisher; #PA1-10008, RRID:AB_1077024; 1:500 in PTwH), TO-PRO™-3 iodide (Thermo Fisher; #T3605; 1:1000 in PTwH.) As secondary antibodies donkey anti-rabbit Alexa Fluor^® 568 (Thermo Fisher; #A10042, RRID:AB_2534017), donkey anti-chicken Alexa Fluor^® 488 (Dianova, Germany; #703-545-155, RRID:AB_2340375), donkey anti-guinea pig Alexa Fluor^® 647 (Dianova; #706-605-148, RRID:AB_10895029) were used, each at dilution of 1:500 in PTwH.

For immunohistochemical analysis, hCOs were fixed with 4% ice-cold paraformaldehyde at 4 °C for 1 h. Fixed hCOs were embedded in Optimal Cutting Temperature (OCT) Compound and sectioned at 20 μm. Sections were blocked for 1 h in PBS containing 0.3% Triton X-100 and 10% normal goat serum. The sections were incubated at 4 °C overnight with primary antibodies in blocking solution, as follows: rabbit anti-SOX2 (1:300; Cell Signaling Technology, Danvers, MA), rabbit anti-TUJ1 (Alexa Fluor^®-488 Conjugate; 1:500; MilliporeSigma, Burlington, MA), chicken anti-MAP2 (1:1000; Abcam, Waltham, MA), rabbit anti-GFAP (1:2000; Dako, Santa Clara, CA), rabbit anti-FOXP2 (1:1000; Abcam), rabbit anti-OLIG2 (1:300; MilliporeSigma), chicken anti-MBP (1:2000; ThermoFisher Scientific, Waltham, MA). All secondary antibodies were ThermoFisher Scientific Alexa Fluor™-conjugated secondary antibodies used at a dilution of 1:500. The sections were imaged with Zeiss Axio Imager.M2.

For immunohistochemical analysis, hCOs were fixed with 4% ice-cold paraformaldehyde (PFA) at 4°C for 1 hr. Fixed hCOs were embedded in Optimal Cutting Temperature (OCT) Compound and sectioned at 20 μm. Sections were blocked for 1 hr in PBS containing 0.3% Triton X-100 and 10% normal goat serum. The sections were incubated at 4°C overnight with primary antibodies in blocking solution, as follows: rabbit anti-SOX2 (1:300; Cell Signaling Technology, Danvers, MA), rabbit anti-TUJ1 (Alexa Fluor^®−488 Conjugate; 1:500; MilliporeSigma, Burlington, MA), chicken anti-MAP2 (1:1000; Abcam, Waltham, MA), rabbit anti-GFAP (1:2000; Dako, Santa Clara, CA), rabbit anti-FOXP2 (1:1000; Abcam), rabbit anti-OLIG2 (1:300; MilliporeSigma), chicken anti-MBP (1:2000; ThermoFisher Scientific, Waltham, MA). All secondary antibodies were ThermoFisher Scientific Alexa Fluor^™-conjugated secondary antibodies used at a dilution of 1:500. The sections were imaged with Zeiss Axio Imager.M2.