Fig 1. MyoF is expressed in HBZ-expressing cells and in HTLV-1-infected T-cells.

(A) Relative MYOF mRNA levels in the indicated HeLa cell lines. The graph shows qRT-PCR results averaged from four independent experiments with values normalized to that of the empty-vector clone, pcDNA (set to 1), for each replicate. Error bars show standard deviations; ** p<0.01, *** p<0.001. (B) MyoF expression in empty vector (pcDNA) and HBZ-HeLa clones. Whole cell extracts (50 μg) were analyzed by Western blot using antibodies against MyoF, HBZ (6xHis) and β-actin. (C) Relative MYOF mRNA levels in resting and activated primary CD4⁺ T-cells and in T-cell lines. The graph shows qRT-PCR results averaged from two independent experiments in which values were normalized to those for the resting CD4⁺ T-cells (set to 1), error bars show standard deviations. Numbers above the bars indicate expression levels relative to the resting CD4⁺ T-cells. HTLV-1-negative lines are T-cell acute lymphoblastic leukemia lines (CEM, HSB-2, Jurkat, Molt-4 and SupT1) and cutaneous T-cell leukemia lines (HUT-78 and Myla); in vitro transformed HTLV-1-positive cells are 1185, C10/MJ, MT-2, MT-4 and SLB-1; ATL-derived cell lines are ATL-2, HUT-102, SP, MT-1 and TL-Om1. aCD4+ is activated CD4⁺ T-cells. BXPC3 is a pancreatic cancer cell line. (D) MyoF expression in T-cell lines. Whole cell extracts (45 μg) were analyzed by Western blot using antibodies against MyoF, Tax, β-actin and HBZ. Endogenous HBZ was enriched by affinity precipitation using GST-KIX (Ponceau S-stained). (E) Tukey boxplots (showing MYOF transcript levels partitioned by healthy donor (HD), asymptomatic carrier (AC), HTLV-1 associated myelopathy (HAM) and (F) adult T-cell leukemia (ATL) from published microarray data: GEO accession numbers GSE29312 [103] and GSE14317 [104] for (E) and (F), respectively; boxplot ends of each whisker are set to 1.5 times the interquartile range above the third quartile and below the first quartile; ** p<0.01, *** p<0.001.