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. 2021 May 10;22(9):5052. doi: 10.3390/ijms22095052

Figure 1.

Figure 1

Figure 1

Crude extract (CE) of C. abbreviata inhibits HIV-1 entry into cells. (A) PBMCs isolated from healthy donor were treated with CE, or the NNRTI inhibitor efavirenz (EFV), the NRTI inhibitor azidothymidine (AZT) or the fusion inhibitor enfuvirtide (T20) for 7 days during infection. HIV-1 infection was assessed by measuring P24 in cell supernatants via ELISA. (B) PBMCs were treated with CE for 2 days. Cell viability was measured by flow cytometry. (C) PBMCs treated with CE were stained with Annexin-V/PI and measured by flow cytometry. Apoptosis level was calculated by counting both early apoptotic cells (Annexin-V+) and late apoptotic cells (PI+) (D,E). CE was tested in a multi-dosing time assay (D) using U373-CD4-CXCR4 cells against pseudotype virus pNL4.3ΔEnvLuc-HXB2 and (E) using U373-CD4-CCR5 cells against pseudotype virus pNL4.3ΔEnvLuc-BAL. CE was either pre-incubated with cells or viruses for 2 h, or incubated with cells and viruses during the 2 h spinoculation infection (normal treatment), or added to the cells after infection (post-incubation). HIV-1 infection was assessed by measuring luciferase in the cell supernatant. Three independent experiments were performed in triplicates.