Fig 2: WNT4 expression in healthy and leukemia-derived cells. A) Relative expression levels of WNT4 were measured by qRT-PCR in PBMCs obtained from healthy volunteers (PBMC) and leukemia-derived cells lines (Jurkat, CEM, HL60, K562, and BJAB). An expression value of the PBMCs of one individual was set as 1. Analysis was calculated using ribosomal Protein L32 (RPL32), ribosomal Protein S18 (RPS18), and beta actin (ACTB) as reference genes. The graphic depicts the means and subsequent standard deviations (SD) obtained with all reference genes. B) Western blot assay showing the presence of WNT4 protein in PBMCs from two healthy individuals and leukemia-derived cells lines. ACTB, beta 2 microglobulin, and a tubulin were used as protein loading controls. C) Box plot graphic showing relative expression levels of WNT4 measured by qRT-PCR normalized to the previously mentioned reference genes. The graph displays median (dark lines), 25–75th percentile (boxes), interquartile ranges (whiskers), and outliers (small, dark circles) from the CD3+ and CD19+ sorted cells of five healthy individuals, cell lines (Jurkat, CEM, HL60, K562, and BJAB), and 11 patients with leukemia. Average values from the PBMCs obtained from the five healthy volunteers were used as controls. Statistical significances are shown between both groups. Experiments were carried out at least twice.

Fig 3: Expression of WNT7A in total peripheral blood cells, Peripheral blood mononuclear cells (PBMC), sorted CD3+ and CD19+ cells obtained from healthy volunteers' samples. (A) Dot blot graphics showing the selected region for sorting (upper left panel), CD3-Fluorescein isothiocyanate (FITC), and CD19-Phycoerythrin (PE) marked cells (upper right panel), isolated CD19+ cells (lower left panel), and isolated CD3+ cells (lower right panel). (B) Figure showing the average of the WNT7A relative expression ratio ± Standard deviation (SD) obtained from the different groups. Normalization was performed by setting CP values of PBMC as 1 and taking GAPDH, RPL32, and RPS18 as reference genes. Experiments were carried out at least twice in all cases.

Fig 4: Cell-surface functionality of CD81 mutants. Huh-7 CD81 KO cells were co-transduced with lentivectors encoding human CD19 and CD81 or empty vector. A, representative flow cytometry histograms. All samples received CD19 lentivector plus the indicated CD81/control vector. Panel i demonstrates CD81 surface expression, and panel ii displays CD81-dependent trafficking of CD19 to the cell surface. B, CD81 expression on CHO cells confers binding on soluble HCV E2. Panel i demonstrates CD81 surface expression, and panel ii displays sE2 binding to transduced CHO cells. C, quantification of sE2 binding expressed relative to WT CD81. An asterisk indicates statistical significance from WT (n = 3, one-way ANOVA, Prism). Error bars indicate standard deviation of the mean.

Fig 5: Colocalization of SRY and cMyb with T-lymphocyte and B-lymphocyte markers in human bone tissue.a Bone tissue of a male patient with osteoporotic fracture showing costaining of SRY with the T-lymphocyte marker CD3 (arrow) in bone marrow (BM) cells. Double-positive cells (arrows) and single-positive cells for SRY or CD3 (arrowhead) are observed within bone marrow cells. b Bone tissue of a male patient with osteoporotic fracture showing costaining of cMyb with the B-lymphocyte marker CD19. Double-positive cells (arrow) and single-positive cells for cMyb or CD19 (arrowhead) are observed within bone marrow (BM) cells. c Bone tissue of a male patient with osteoporotic fracture showing costaining of cMyb with the T-lymphocyte marker CD3 (arrow) in bone marrow (BM) cells. Double-positive cells (arrows) are observed within bone marrow cells. Single-positive cells for cMyb (arrowhead) are observed within bone marrow cells, on the trabecular bone (TB) surface, and in a few osteocytes. d Bone tissue of female patients with osteoporotic fracture showing negative staining for SRY and similar patterns of cMyb staining as in the male osteoporotic fracture tissue. Scale bars: 50 µm