Ab25195

CD4-VioGreen (130-106-655, Miltenyi Biotec), Anti-CCR10-PE (clone: REA326, 130-104-822, Miltenyi Biotec), CD183 (CXCR3)-PE-Vio770 (clone: REA326, 130-104-822, Miltenyi Biotec), CD194 (CCR4)-APC (clone: REA279, 130-103-813, Miltenyi Biotec), CD196 (CCR6)- PE-Vivo-615 (clone: REA190, 130-107-142, Miltenyi Biotec), Propidium iodide (130-093-233, Miltenyi Biotec), NESTIN rabbit-anti-mouse (ab7659, Abcam), SCA1 rat-anti-mouse (ab25195, Abcam), PDGFRα goat-anti-mouse (AF1062, R&D Systems), Alexa Flour 405 donkey-anti-Rabbit (ab175651, Abcam), Alexa Flour 488 donkey-anti-goat (A-11055, Thermo Fisher Scientific), Alexa Flour 594 donkey-anti-rat (A-21209, Thermo Fisher Scientific), PerCP-CyTM 5.5 CD45.2 conjugated mouse-anti-mouse (552950, BD Biosciences), ICAM1 rabbit-anti-human (ab53013, Abcam), VCAM1 rabbit-anti-human (ab134047, Abcam), IL-1β rabbit-anti-human (ab2105, Abcam), TNF-α rabbit-anti-human (ab9635, Abcam), actin rabbit-anti-human (A2066, Sigma-Aldrich), rabbit anti goat (81–1620, Invitrogen), goat anti rabbit (65–6120,Invitrogen), CD3 rabbit-anti mouse (AD452, Dako), and B220 rat-anti-mouse (MAB1217, R&D Systems).

Briefly, slices (or cells) were incubated for 1 h with 10 % normal donkey serum in PBS 0.1 M (supplemented with 0.3 % Triton X-100 for intracellular antigens). For specific immunofluorescent staining, anti-nestin (1:300, NB100-1604, Novus Biologicals, Littleton, CO, USA), anti-Sox2 (1:250; sc-17320, Santa Cruz, Dallas, Texas, USA), anti p75NTR (1:200, AB1554, Millipore, Darmstadt, Germany), anti-Sca-1 (1:100, ab25195, Abcam, Cambridge, United Kingdom), anti-arginase 1 (1:100, 610708 BD Biosciences, Franklin Lakes, NJ, USA), anti-Iba1 (1:800, 019-19741, Wako Chemicals, Richmond, VA, USA), anti-GFAP (1:1000, 20334, Dako, Glostrup, Denmark), anti-laminin (1:200, L9393, Sigma-Aldrich, Saint-Louis, MO, USA) were diluted in PBS 0.1 M overnight at 4 °C. After PBS washes, brain sections were incubated at room temperature with fluorescein and rhodamine red X-conjugated secondary antibodies (1:500; Jackson Immunoresearch Laboratories, Westgrove, PA, USA) or with peroxidase-coupled secondary antibodies (1:500, Dako, Glostrup, Denmark) and diaminobenzidine revelation. Nuclei were counterstained with Hoescht for fluorescent staining. Image acquisition and analysis were performed using a Zeiss AxioImager Z1 epifluorescent microscope coupled with FluoView (Olympus), and Olympus AX-70 microscope coupled with AnalySIS software (Olympus).