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2 protocols using fitc isotype control

1

Flow Cytometry Analysis of TDSC Markers

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TDSCs were incubated with fluorescent primary antibodies on ice for 60 min, washed three times with phosphate-buffered saline (PBS), and analyzed by flow cytometry. The following fluorescent primary antibodies were used: FITC anti-rat CD29, FITC anti-rat CD44, PE anti-rat CD45, and PE anti-rat CD90 (BioLegend, San Diego, CA, United States). FITC isotype control and PE iso control were added to the NC samples (BioLegend, San Diego, CA, United States).
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2

HIV-1 BaL Infection Pathway Analysis

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HIV-1 BaL was obtained from the NIH AIDS Research and Reference Reagent Program, National Institute of Allergy and Infectious Diseases, NIH. The HIV-1 BaL strain has been shown to efficiently infect, via the CCR5 co-receptor, and replicate within CD4+ T-cells49 (link). Argonaute-1, p-NFκB, NFκB, p-ERK, p-Src, p-Akt, NFAT1, p-CREB, CREB, Oct-1, Ubiquitin, and β-Actin antibodies were obtained from Cell Signaling Technology (Danvers, MA). GW182, D1DR, D2DR, D3DR, D4DR, Sigma-1 Receptor, and GAPDH antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). FITC-conjugated p24 GAG (6604665) antibody was obtained from Beckman Coulter, Inc. (Brea, CA). PE-conjugated CD45RO, PE-conjugated HLA-DR, FITC-conjugated CD45RA, FITC-conjugated CD25, FITC-conjugated CD69, PE isotype control, and FITC isotype control antibodies were purchased from Biolegend (San Diego, CA). p-NFAT1 antibody was obtained from Thermo Fisher Scientific (Waltham, MA). Fluo-4, AM was purchased from Invitrogen (Carlsbad, CA). Methamphetamine hydrochloride and sigma-1 receptor inhibitor (BD1047) were purchased from Sigma Aldrich (St. Louis, MO).
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