Mip 1α

RPMI-1640 medium, penicillin and streptomycin, PD098059 (ERK inhibitor), PP2 (LYN inhibitor), Brefeldin A (Golgi blocker) and TRI Reagent were from Sigma (St Louis, MO, USA). Fetal calf serum (FCS) was purchased from Gibco BRL (Grand Island, NY, USA) and ELISA Assay Kit of mouse IL-10, IL-12, TNF-α, IFN-γ, MIP-1α, MIP-1β and Rantes were from BD and TGF-β was from eBioscience. dNTPs, RevertAidTM M-MuLV Reverse Transcriptase, oligo dT, RNase inhibitor and other chemicals required for cDNA synthesis were from Fermentas (USA). Anti-phospho-H3 and Anti-acetyl-H3 Abs were obtained from Abcam and chromatin immunoprecipitation (ChIP) assay kits were purchased from Millipore (Bedford, MA, USA). GAPDH, phosphorylated and dephosphorylated form of Lyn and ERK-1/2 antibodies, CCR5 antibody were obtained from Santa Cruz Biotechnology (San Jose, CA, USA).

Similar to the evaluation of NK cell activation, supernatants of co-cultured cells were collected, however they were obtained from independent wells and stored at − 80 °C until they were used. Supernatants were thawed at 4 °C right before running the CBA assay. The panel for the CBA flex set included: TNF-α, Granzyme, IFN-γ, MIP-1α and RANTES (BD Biosciences, San Jose, CA, USA). The CBA assay was done according to the manufacturer´s instructions. The beads complex was acquired using LS Fortessa (BD Biosciences, San Jose, CA, USA). Obtained data were normalized with NK cells co-cultured with uninfected CD4⁺ T cells and analyzed using FlowJo version 10.5.3 (FlowJo, LLC, Oregon, USA).