Fig. 1

Characterization of mature microglia and BO morphology in response to pro-inflammatory stimuli. A H&E staining of 4-month-old BO. The image shows typical BO morphology with classical rosette structures. The magnified section indicates the location of neuro stem cells (NSCs) and mature neurons (Neu) within the rosette structure. The immunofluorescence of the rosette structure confirm the presence of NSCs (Nestin) and Neu (MAP2) B Characterization of one BO by obtained by flow cytometry analysis, identifying different populations: 5.48% GFAP + reactive astrocytes (Ast), 69.3% Tuj-1 + neurons (Neu), 0% Iba-1 + /P2RY12 + microglia (Mic), and 9.7% PAX6 + /SOX2 + neuro NSCs. C p-NF-KB levels in one BO (n = 1) stimulated with TNFa 40 ng/ml + 100ng/ml IFN in presence of 100 nM calyculin versus a BO treated with vehicle (DMSO) for 1 h, obtained by flow cytometry analysis. The dot plot shows the changes of p-NF-κB in the different BOs population characterized by CNS markers: GFAP + (reactive astrocyte), TUJ-1 (neurons) and SOX2 + (neuro NSCs). D Characterization of fully differentiated microglia (HPCs -derived microglia) after 11 days shows typical microglial morphology. Representative flow cytometry dot plot showing the expression of classical microglial markers: CD11b (82.7%), CD45 (low), Iba-1 (80.8%), P2RY12 (low), and TREM2 (22%). E Analysis of Gag expression in HIV-1 infected microglia by immunofluorescence 24 h post-infection (top images). The arrows in the images indicate syncytia structures with high Gag protein expression, characteristic of HIV-1 infection (n = 1). F Quantification of levels of p24 detected in the supernatant of HIV-1 infected microglia by ELISA over 9 days of incubation (graph) (n = 1) G Quantification of pro-inflammatory cytokine levels associated with HIV-1-NCI and IRIS in the supernatant of three different microglia batches at day 3 post-infection (n = 3)