Fig 5. MyoF enhances HTLV-1 infection.

(A) The flow diagram shows the co-culture/infection assay procedure using CHOK1-Luc or Jurkat-pminLUC-vCRE cells as target cells. (B) Inhibition of MyoF reduces HTLV-1 infection. ATL-2 cells were treated with DMSO or WJ460 (200nM or 1μM) prior to co-culture with CHOK1-Luc cells. (C) SLB-1 cells were treated with DMSO or 1 μM WJ460 prior to co-culture with Jurkat-pminLUC-vCRE cells. (D) C8166/45 and SLB-1 cells (both high Tax expression) and TL-Om1 cells (no Tax expression) were co-cultured with Jurkat-pminLUC-vCRE cells. (E) An HTLV-1-immortalized primary human T-cell clone (C13) was treated with DMSO or 1 μM WJ460 prior to co-culture with Jurkat-pminLUC-vCRE cells. Graphs (B), (C) and (D) show luciferase assay results averaged from at least three replicates for each condition of a single experiment and are representative of three independent experiments; * p<0.05, *** p<0.001. Graph (E) shows luciferase assay results average from three replicates for each condition of a single experiment and is representative of two independent experiments; ** p<0.01. (F) Knockdown of MyoF expression reduces HTLV-1 infection. MyoF expression in control (GFP) and MyoF knockdown ATL-2 and SLB-1 cells. Whole cell extracts (50 μg) were analyzed by Western blot using antibodies against MyoF and β-actin. (G) SLB-1 cells stably expressing an shRNA targeting GFP (negative control) or MYOF mRNA were co-cultured with CHOK1-Luc cells. (H) SLB-1 cells stably expressing an shRNA targeting GFP (negative control) or MYOF mRNA were co-cultured with Jurkat-pminLUC-vCRE cells. (I) ATL-2 cells stably expressing an shRNA targeting GFP (negative control) or MYOF mRNA were co-cultured with Jurkat-pminLUC-vCRE cells. Graphs show luciferase assay results averaged from at least three replicates for each infection condition of a single experiment and is representative of at least three independent experiments; ** p<0.01, *** p<0.001.