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. 2009 Apr 28;7(4):e1000095. doi: 10.1371/journal.pbio.1000095

Figure 2. Extracts from HL60 Cells Stably Transfected with Retrocyclin Constructs Are Active against HIV-1 Infection.

Figure 2

(A) TZM-bl cells were treated with extracts or peptide as indicated in the figure and infected with HIV-1 BaL (6.5 ng/ml p24) for 24 h. Infection was measured as percent luciferase activity compared to cells treated with control cell extract (average relative luciferase units [RLUs] of control HL60 extract = 178,200).

(B, C) PM1 cells and PBMCs were treated with extracts or peptide as indicated and infected with HIV-1 BaL (2 ng/ml p24) and cultured for 5–9 d. Bars represent percent BaL HIV-1 levels in the supernatants collected on days 5 (B) and 9 (C). The amount of p24 in PM1 cells treated with control extract = 76.85 ng/ml and in PBMCs treated with control extracts = 55.99 ng/ml.

(D) TZM-bl cells were treated with immunopurified (IP) extracts or peptides as indicated and infected with BaL HIV-1 (p24 = 2 ng/ml) for 24 h. Infection was quantified as percent luciferase activity compared to cells treated with control HL60 cell IP extracts (average RLU = 764,460). Error bars represent standard error of the mean (SEM). n = 3–4; #, p < 0.004; *, p < 0.002; **, p <0.0005.

(E) Cellular viability of TZM-bl cells treated with HL60 acid extracts as indicated was determined by measuring the reduction of MTT after 24h (n = 3). Bars represent percent viability as compared to vehicle control and error bars represent SEM.

(F) Cell death was monitored in PBMCs treated with the acid extracts by a trypan blue exclusion assay on day 9 (n = 1).