s5
Homo sapiens
Protocols: Monocytes were isolated from the peripheral blood of healthy donors by Ficoll-Paque density centrifugation followed by immunomagnetic selection using CD14+ microbeads (MACS monocyte isolation kit, Miltenyi Biotec) according to manufacturer's instructions. MDMs were treated with anti-CCL2 or control Ab for 4 h and then infected with 3,000 tissue culture infectious dose (TCID50) per well of the CCR5-dependent HIV-1BaL strain pelleted virus (Advanced Biotechnologies), corresponding to a MOI ≈ 0.03. After 2 h, cells were washed and maintained in complete medium either in the presence or in the absence of anti-CCL2 or control Ab Total RNA samples were isolated with the RNeasy Mini kit (Qiagen) following the manufacturer's instructions. RNA concentration and integrity were analyzed by a NanoDrop 2000 (Thermo Scientific) and a TapeStation (Agilent). Total RNA was subjected to poly (A) selection followed by reverse transcription