Journal: bioRxiv
Article Title: Neuroinflammation and EIF2 signaling persist in an HiPSC tri-culture model of HIV infection despite antiretroviral treatment
doi: 10.1101/779025
Figure Lengend Snippet: Generation and characterization of iMg (a) Timeline for iMg differentiation and HIV infection. (b) Immunostaining for microglia specific markers: CX3CR1, TMEM119, IBA1, and P2RY12. All cultures were stained for DAPI. (c) Bulk RNAseq normalized read counts of specific microglia markers from iMg and MDMs. n=4 iMg, n=3 MDMs. Benjamini-Hochberg, FDR 0.01, **p
Article Snippet: Antibodies used for immunofluorescence: Chicken anti-MAP2 (Abcam ab5392, RRID:AB_2138153, 1:500); Mouse anti-PSD-95 clone K28/43 (NeuroMab 75-028, RRID:AB_2292909, 1:500); Mouse anti-Synaptophysin clone SY38 (Millipore MAB5258-20UG, RRID:AB_95185, 1:250); Mouse anti-Nestin clone 10C2 (Millipore MAB5326, RRID:AB_11211837, 1:200); Rabbit anti-Thrombospondin-1 (Abcam ab85762, RRID:AB_10674322, 1:250); Mouse anti-Glutamine Synthetase (Millipore MAB302, RRID:AB_2110656, 1:500); Rabbit anti-SOX9 (Abcam ab185230, RRID:AB_2715497, 1:250); Rabbit anti-CX3CR1 (Abcam ab8021, RRID:AB_306203, 1:500); Rabbit anti-TMEM119 (Abcam ab185333, RRID:AB_2687894, 1:200); Rabbit anti-IBA1 (Wako 019-19741, RRID:AB_839504, 1:500); Rabbit anti-P2RY12 (Alomone Labs APR-020, RRID:AB_11121048, 1:100); Rat anti-LAMP1 (Abcam ab25245, RRID:AB_449893, 1:500); Mouse anti-HIV1 p24 [39/5.4A] (Abcam ab9071, RRID:AB_306981, 1:500); Rabbit anti-Human Nanog (Cell Signaling Technology 3580, RRID:AB_2150399, 1:800); Mouse anti-NCAM Clone 2-2b (Millipore MAB5324, RRID:AB_95211, 1:250); Rabbit anti-OCT-4 (Cell Signaling Technology 2750, RRID:AB_823583, 1:200); Rabbit anti-SOX2 (Millipore AB5603, RRID:AB_2286686, 1:100); Rabbit anti-CCR5 (Thermo Fisher Scientific pa5-29011, RRID:AB_2546487, 1:500); Mouse anti-GLAST (Miltenyi Biotec 130-095-822, RRID:AB_10829302, 1:50); Mouse anti-GFAP (Sigma-Aldrich SAB1405864, RRID:AB_10739114, 1:10,000); Mouse anti-Beta-Actin (Cell Signaling Technology 3700, RRID:AB_2242334, 1:10,000); Rabbit anti-EAAC1 (Santa Cruz Biotechnology sc-25658, RRID:AB_2190727, 1:50); Rabbit GLT-1 (Jeff Rothstein Lab 1:5,000); Goat anti-mouse IgG (H+L) Alexa Fluor 488 (Thermo Fisher Scientific A-11029, RRID:AB_138404, 1:500); Goat anti-rabbit IgG (H+L) Alexa Fluor 488 (Thermo Fisher Scientific A-11034, RRID:AB_2576217, 1:500); Goat anti-mouse IgG (H+L) Alexa Fluor 568 (Thermo Fisher Scientific A-11004, RRID:AB_2534072, 1:500); Goat anti-rabbit IgG (H+L) Alexa Fluor 568 (Thermo Fisher Scientific A-11036, RRID:AB_10563566, 1:500); Goat anti-rat IgG (H+L) Alexa Fluor 568 (Molecular Probes A-11077, RRID:AB_141874, 1:500); Goat anti-rat IgG (H+L) Alexa Fluor 680 (Molecular Probes A-21096, RRID:AB_141554, 1:500); Goat anti-Chicken IgY (H+L) DyLight 680 (Thermo Fisher Scientific SA5-10074, RRID:AB_2556654, 1:500).
Techniques: Infection, Immunostaining, Staining