Auranofin potently upregulates hepcidin expression in Huh7 cells. aHAMP1 mRNA was measured in Huh7 cells treated for 12 h with 5 μM of the indicated drugs (see Table S1 for details); all 106 drugs caused a significant change in HAMP1 mRNA compared to control-treated cells. bHAMP1 mRNA levels were measured in Huh7 cells treated with dihydro-ergotamine Mesylate (1 μM) or AUR (1 μM) for 12 h. cHAMP1 mRNA was measured in Huh7 cells treated for 12 h with the indicated concentration of AUR or BMP6(50 ng/ml). dHAMP1 mRNA was measured in Huh7 cells treated with 0.5 μM AUR for the indicated time. e Cell viability assay was performed in Huh7 cells treated with the indicated concentration of AUR for either 12 or 18 h. f Huh7 cells were co-transfected with pGL3-HAMP1 and the Renilla reporter construct; 36 h after transfection, the cells were treated with AUR (0.5 μM) or 50 ng/ml BMP6 for 18 h, after which luciferase activity was measured. The mRNA levels in panels a, b and d were normalized to β-ACTIN and are expressed relative to the mean control value. The cell line based in vitro experiments were repeated at least three independent times. Error bars indicate the SEM. The data in a and e were analyzed using the Student’s t-test (*p < 0.05). The data in b–d were analyzed using a one-way ANOVA with Tukey’s post hoc test; groups labeled without a common letter were significantly different (p < 0.05)
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机译：Auranofin在HuH7细胞中高效地提出了Hepcidin表达。在处理12小时的HUH7细胞中测量AHAMP1 mRNA，用5μm表示的药物（有关详细信息，见表S1）;与对照处理处理的细胞相比，所有106种药物导致Hamp1 mRNA的显着变化。在用二氢甲丙胺甲磺酸根（1μm）或α（1μm）处理的HUH7细胞中测量BHAMP1 mRNA水平。12小时。将CHAMP1 mRNA在处理12小时的HUH7细胞中测量，浓度或BMP6（50ng / mL）浓度。在用0.5μmAur处理的HuH7细胞中测量DHAMP1 mRNA，用于指示时间。 E细胞活力测定在用指定的浓度为12或18小时处理的HUH7细胞中进行。将F HUH7细胞与PGL3-HAMP1和Renilla报告构建体共转染;转染后36小时，用Aur（0.5μm）或50ng / ml BMP6处理细胞18小时，然后测量荧光素酶活性。板A，B和D中的mRNA水平标准化为β-肌动蛋白，并且相对于平均控制值表达。基于体外实验的细胞系重复至少三次。误差条表示SEM。使用学生的T检验分析A和E中的数据（* P <0.05）。使用单向ANOVA分析B-D中的数据，具有Tukey Hoc测试;没有常见字母标记的组显着不同（P <0.05）
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