Figure 1
Figure 1. T-plastin mRNA and protein expression in lymphocytes from patients with SS, MF, various hemopathies or undetermined erythroderma and from healthy donors. Total mRNA was extracted from blood lymphocytes of patients with SS (n = 94), MF (n = 34), hemopathies, undetermined erythroderma and scleroderma (n = 38), and of healthy donors (n = 78), purified and used for qRT-PCR (A-D) or multiplex standard PCR (C). Results from qRT-PCR (A-D) are obtained as ΔCt values expressed in arbitrary units (AU) and they are presented as mean ± SEM in panel A, as individual data in panels B through D, and in panel B, with a dark line showing mean values for each set of data. Results in panel C are from multiplex standard PCR of PBLs from 8 SS patients, 1 MF patient, 1 healthy volunteer and for purified CD4+Vβ2+ and CD4+Vβ2− cells from SS patient no. 4 as well as purified CD4+ cells from 1 healthy donor. CD4+ cells were isolated by magnetic-activated cell sorting using the CD4 cell isolation kit (Miltenyi Biotec) and further Vβ2+ isolation using anti-TCR Vβ2 (Clone MPB2D5) from Beckman Coulter and magnetic dynabeads goat anti–mouse IgG. For each CTCL patient are given percentage of Sézary cells (SS%), detection of T cell clone by Vγ-Jγ PCR presented as positive (+) or negative (−), percentage of positive Vβ/CD4 subset and Vβ type obtained by immunophenotyping using flow cytometry (except immunoscope analysis for Vβ6.1). nd indicates not determined. In panel D, under T-plastin mRNA expression obtained by qRT-PCR is shown respective T-plastin protein expression from total extract of PBLs from SS patients (SS, n = 17), 1 MF patient, 1 healthy donor, and HuT-78 cells, as assessed by Western blotting. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) expression is presented as a loading control.

T-plastin mRNA and protein expression in lymphocytes from patients with SS, MF, various hemopathies or undetermined erythroderma and from healthy donors. Total mRNA was extracted from blood lymphocytes of patients with SS (n = 94), MF (n = 34), hemopathies, undetermined erythroderma and scleroderma (n = 38), and of healthy donors (n = 78), purified and used for qRT-PCR (A-D) or multiplex standard PCR (C). Results from qRT-PCR (A-D) are obtained as ΔCt values expressed in arbitrary units (AU) and they are presented as mean ± SEM in panel A, as individual data in panels B through D, and in panel B, with a dark line showing mean values for each set of data. Results in panel C are from multiplex standard PCR of PBLs from 8 SS patients, 1 MF patient, 1 healthy volunteer and for purified CD4+Vβ2+ and CD4+Vβ2 cells from SS patient no. 4 as well as purified CD4+ cells from 1 healthy donor. CD4+ cells were isolated by magnetic-activated cell sorting using the CD4 cell isolation kit (Miltenyi Biotec) and further Vβ2+ isolation using anti-TCR Vβ2 (Clone MPB2D5) from Beckman Coulter and magnetic dynabeads goat anti–mouse IgG. For each CTCL patient are given percentage of Sézary cells (SS%), detection of T cell clone by Vγ-Jγ PCR presented as positive (+) or negative (−), percentage of positive Vβ/CD4 subset and Vβ type obtained by immunophenotyping using flow cytometry (except immunoscope analysis for Vβ6.1). nd indicates not determined. In panel D, under T-plastin mRNA expression obtained by qRT-PCR is shown respective T-plastin protein expression from total extract of PBLs from SS patients (SS, n = 17), 1 MF patient, 1 healthy donor, and HuT-78 cells, as assessed by Western blotting. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) expression is presented as a loading control.

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