Journal: Communications Biology

Article Title: NHC-gold compounds mediate immune suppression through induction of AHR-TGFβ1 signalling in vitro and in scurfy mice

doi: 10.1038/s42003-019-0716-8

Figure Lengend Snippet: a Heatmap of CYPs expression changes upon MC3 treatment for 1 h and 24 h from DNA microarray data (values are log of fold expression change versus DMF treatment). Expression profile of all members of the CYP 450 family and genes related to AHR-NRF2 signaling pathway. b Expression of CYPs and AHR-NRF2 related genes after MC3 treatment analyzed by RT-qPCR ( n ≥ 6). c AHR antagonist resveratrol (Res) significantly represses CYP1A1 expression in HepG2 cells treated with MC3 1 µ m or TCDD 10 n m for 1 h. Data were normalized to the TCDD-induced CYP1A1 expression ( n = 9). d Comparison of CYP1A1 expression in HepG2 AHR KD and CRISPR/CAS control cells, indicated as Ctrl. ( n = 9). e Comparison of CYP1A1 and AHR expression in HepG2 control and AHR KD cells treated with TCDD 10 n m or MC3 1 µ m for 24 h. One-way ANOVA t test was performed. * p < 0.05, ** p < 0.01, *** p < 0.001;. lower and upper ends of the bars indicate the minimum and maximum values, respectively, and the centre represents the median. Error bars ± SD. The source data for 1 b – d are provided as Supplementary Data .

Article Snippet: The basic technical specifications of this system have been described previously and are outlined in Supplementary Fig. . We used hepatocellular carcinoma cell line HepG2 as a metabolite generator, which expresses the majority of human cytochromes P450 (CYPs) and is widely used in various microfluidic systems , , , .

Techniques: Expressing, Microarray, Quantitative RT-PCR, Comparison, CRISPR, Control

Journal: Communications Biology

Article Title: NHC-gold compounds mediate immune suppression through induction of AHR-TGFβ1 signalling in vitro and in scurfy mice

doi: 10.1038/s42003-019-0716-8

Figure Lengend Snippet: a Functional annotation analysis of DNA microarray data of RNA collected from HepG2 cells treated with 1 µ m MC3 for 1 h and 24 h. b – e Inhibitory effect on primary CD4 + T-cell activation. Resting CD4 + T cells were left untreated or incubated overnight with 0.5 µ m MC3, 0.5 µ m auranofin or 10 n m TCDD. Cells were then activated through CD3-CD28 stimulation and assayed for: b ACTIN remodelling (formation of ACTIN rings) with the schematic representation of the treatment (Scale bar: 10 µ m ); c CD25 expression; d IL-2 production or e CD38 expression. For b , n = 3 donors, for c – e , n = 5 donors. Data were analyzed by one-way ANOVA or Friedman test. An appropriate transformation (Logit) was employed to restore normality when appropriate. One-way ANOVA t test was performed. * p < 0.05, ** p < 0.01, *** p < 0.001; lower and upper ends of the bars indicate the minimum and maximum values, respectively, and the centre represents the median. Error bars ± SD. The source data for 2 c – e are provided as Supplementary Data .

Article Snippet: The basic technical specifications of this system have been described previously and are outlined in Supplementary Fig. . We used hepatocellular carcinoma cell line HepG2 as a metabolite generator, which expresses the majority of human cytochromes P450 (CYPs) and is widely used in various microfluidic systems , , , .

Techniques: Functional Assay, Microarray, Activation Assay, Incubation, Expressing, Transformation Assay