Fig 2: Effect of curcumin on the histone H3 acetylation levels of TIMP1 promoters in spontaneous hypertensive rats. (A) PCR-amplified genomic DNA obtained following ChIP with an anti-Ac-H3 antibody. (B) PCR-amplified genomic DNA obtained following ChIP with an anti-HDAC1 antibody. *P<0.05 vs. non-treated, **P<0.01 vs. non-treated, n=6 in each group. ChIP, chromatin immunoprecipitation; IP, immunoprecipitation; PCR, polymerase chain reaction; anti-Ac-H3, anti-acetylated histone H3.

Fig 3: Curcumin inhibited invasion of PANC-1 cells alone or combined with gemcitabine or docetaxel. (A-C) Matrigel invasion assay was conducted to observe the invasion capacity of PANC-1 cells under different drug regimens. (D-F) Quantitative analysis of invasive PANC-1 cells when added with or without curcumin (*P<0.05). (G-I) Expression of MMP2, MMP9, TIMP1 and TIMP2 in PANC-1 cells at indicated treatment were measured by western blot analysis.

Fig 4: ECM remodeling mechanism in hyperglycemia is distinct from drug-induced injury A. Schematic of the experimental strategy for hyperglycemia- and drug-induced disease treatments. B. Immunofluorescence microscopy images of podocyte foot process morphology (top and middle row panels: DAPI, blue; nephrin, red) and actin cytoskeleton (bottom row panel: DAPI, blue; F-actin, gray) via hyperglycemia- and drug-induced injury. White arrowheads indicate primary and secondary foot processes, yellow arrowheads indicate foot process retraction, dashed lines denote cell body perimeter C. Podocyte viability before and after hyperglycemia- and drug-induced podocyte injury; n = 3. D. Immunofluorescence images of podocytes (DAPI, cyan; Nephrin, magenta) collagen (IV)α1 (magenta) deposition in hyperglycemia- and drug-induced injury. E. Western blot analysis of embryonic isoform collagen (IV)α1 following nine days of injury. F. Densitometric analysis of collagen (IV)α1 after nine days of injury.; n = 3. G. Gelatinase (MMP-2 and MMP-9) activity normalized to total protein; n = 3 color matched biological replicates. H. Western blot of TIMP-1 expression after 9 days of injury. I. Densitometric analysis of TIMP-1 levels after 9 days of injury; n = 3 color-matched biological replicates for Adriamycin, n = 4 color-matched biological replicates for all other conditions. B, D Scale bars, 100 µm. C, F, G, I *p < 0.05; **p < 0.01. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Fig 5: NRF3 impairs breast cancer cell invasion through the AKT/ID3 axis. (A) .The most ten downregulated genes under NRF3 overexpression obtained from the data of RNA seq, relative expression level, normalized by log10 (FPKM). (B) qRT-PCR was used to validate the RNA seq data, relative expression level, normalized by GADPH. (C) Western blot for ID3, AKT, p-AKT, NF-κB p65, and p-NF-κB p65 protein levels in MDA-MB-231 cells overexpressed with NRF3 or not. β-actin was as a loading control. (D) Western blot (left panel) for ID3, AKT and p-AKT protein levels in MDA-MB-231 cells treated with LY294002 or not. β-actin was as a loading control. (E) Western blot for ID3, E-cadherin and MMP-2 protein levels in MDA-MB-231 cells over-expressed with ID3 or not. β-actin was used as a loading control. (F) Cell invasion assays for MDA-MB-231 cells with overexpressed NRF3, ID3 alone or in combination by transwell with matrigel. Representative images at 40× (left panel), total view for whole well (middle panel) and the quantification for cell number invaded per view (right panel). Error bar indicated SD (*P<0.05 and **P<0.01). Three independent experiments were performed for all assays.Abbreviations: NRF3, nuclear factor, Erythroid 2 Like 3; AKT, AKT serine/threonine Kinase; ID3, inhibitor of DNA binding 3; FPKM, Reads Per Kilobase of exon model per Million mapped reads; MMP, matrix metallopeptidase; TIMP, TIMP metallopeptidase inhibitor.