Fig 1: Serum and tissues copper concentrations, redox balance and MG-H1 levels in ND or HFD mice. a Representative photomicrographs of Hematoxylin and Eosin staining in liver sections from representative liver tissues of male mice fed with ND or HFD. Original magnification ×10. b Copper concentration in serum and liver samples of ND and HFD mice. c Transcriptional levels of CTR1, CTR2, ATP7B, COX17, CCS and ATOX1 genes. d Levels of reduced GSH expressed by tGSH/GSSG ratio and d MG-H1 (methyl-glyoxal-hydro-imidazolone) protein adducts. Values are expressed as means ± SD and data were analyzed by a t-Test analysis (*P < 0.05; **P < 0.01; ***P < 0.001)
Fig 2: CTR1 silencing(A) Relative cell viability evaluated by MTS assay. Results derived from three independent experiments and are represented as mean ± SD, respect to un-transfected cells. (B) Relative mRNA expression of MYC, CTR1 and CTR2 in scramble and siCTR1 transfected cells, in presence or absence of copper treatment (35 µM CuSO4). Values are expressed as mean ± SD (*P < 0.05, **P < 0.01, and ***P < 0.001, respect to un-transfected cells; n = 3). (C) Representative western blot (Top) and the relative densitomentric analysis (Bottom) of MYC, CTR1 and CTR2. Values are expressed as mean ± SD (*P < 0.05, **P < 0.01, and ***P < 0.001, respect to un-transfected cells; n = 3).
Fig 3: Effects of Ole Cu-transporter proteins in ND and HFD mice. RT-PCR, Western blot analysis and relative densitometry of CTR1 (a) and CTR2 (b) proteins. Values are expressed as fold mean ± SD. Data were analyzed by a factorial ANOVA with post-hoc Fisher’s tests for multiple comparison (*P < 0.05, **P < 0.01; ***P < 0.001 vs. ND mice; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. HFD mice; §P < 0.05, §§P < 0.01 vs. male mice)
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