MMP

Proliferation indices of SU-pcGBM2 cells exposed to plain medium (aCSF) or active conditioned medium generated in the presence or absence of pan-MMP inhibitor (BAT).

GM6001 inhibits MMP-7 and −10 mRNA expressions in Aurora-A stable cells in HNC. The mRNA expression levels of MMP-7 and −10 were examined by Q-RT-PCR in FaDu/vehicle, and FaDu/Aurora-A transfectants treated with DMSO or GM6001 in a dose-dependent manner. Statistical analysis: **p < 0.01, ***p < 0.001.

The matrix metalloproteinase 9(MMP9) inhibitor(SB-3CT) attenuated angiotensin II (Ang II)-induced cardiomyocyte hypertrophy in vitro. (A) Representative immunofluorescent (α-actinin) images of neonatal rat cardiomyocytes (NRCMs) in the presence or absence of MMP9 inhibitor that were administered PBS or Ang II for 48 h. Bar=200 μm. (B) Statistical results for the cell surface area (CSA) of NRCMs. (C-E) The relative mRNA expression levels of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC) in NRCMs in the presence or absence of MMP9 inhibitor that were administered PBS or Ang II for 48 h. The intensities were normalized to the levels of GAPDH mRNA with the value in the PBS group defined as 1. * P<0.05 versus the control group, # P<0.05 versus the Ang II group. n=6 samples per experimental group

(E, F) Uev1A mediates negative feedback of NF-κB signaling pathway is dependent on TACE activity. Forty-eight hours after transfection, the cells were pre-treated with BB-2516 (20 μM) for 1 h, followed by TNFα (40 ng/ml) treatment for 2 h. Cell was fixed and stained with antibodies against HA and p65. The same cells were also stained with DAPI to reveal nuclei. White arrows indicate UEV1A-overexpressed cells. (F) Quantitative analysis of the experiments shown in (E). Percentage of cells displaying nuclear p65 is shown for UEV1A- overexpressed or vector-transfected cells. Each sample contains about 1000 cells. The significance of differences was assessed by two-tailed Student's test (***P < 0.001).

NOB modulates Cps1 mRNA and protein expression. a Total protein extracts were prepared from liver samples collected from the four diet/treatment groups of wild-type mice at the indicated circadian time points (n = 3). Western blotting analysis was performed using anti-CPS1 antibody. RC regular chow, HFD high-fat diet, Veh vehicle, NOB Nobiletin. The results shown are representative of three independent experiments. See Additional file 1: Figure S1A for quantitative analysis. b Immunohistochemical staining of CPS1 in liver sections from mice with the indicated diet and treatment at ZT2. c Real-time RT-PCR analysis of Cps1 in liver samples collected as in (a). Data are presented as mean ± SEM (n = 3). Two-way ANOVA with Bonferroni post-hoc tests shows significant statistical differences between HFD.Veh and other three groups (p < 0.0001). d Western blotting analysis of protein lysates of liver samples collected at ZT 6 and 18 from mice with the indicated diet and treatment (n = 3). HPD indicates high-protein diet. The images shown to the left are representative of three independent experiments. Quantitation of Western blots was carried out and the results, presented as mean ± SEM, are shown in the lower panel. Two-way ANOVA with Bonferroni post-hoc tests, RC vs. HPD, ***p < 0.001. e Real-time qPCR analysis was carried out using total RNAs extracted from the liver samples described in (d). The results are presented as mean ± SEM. Two-way ANOVA with Bonferroni post-hoc tests, RC vs. HPD, *p < 0.05.

(B) Florescent images of EpEX (green) and EpICD (red) in PK-15 cells, which were transfected by pSIN-fEpCAM-mCherry and treated with or without TAPI-1/DAPT. Nuclei were stained by Hoechst 33342 (blue). Scale bar, 25 μm.

Proliferation indices of SU-pcGBM2 cells exposed to plain medium (aCSF) or active conditioned medium generated in the presence or absence of pan-MMP inhibitor (BAT).

GM6001 inhibits MMP-7 and −10 mRNA expressions in Aurora-A stable cells in HNC. The mRNA expression levels of MMP-7 and −10 were examined by Q-RT-PCR in FaDu/vehicle, and FaDu/Aurora-A transfectants treated with DMSO or GM6001 in a dose-dependent manner. Statistical analysis: **p < 0.01, ***p < 0.001.

The matrix metalloproteinase 9(MMP9) inhibitor(SB-3CT) attenuated angiotensin II (Ang II)-induced cardiomyocyte hypertrophy in vitro. (A) Representative immunofluorescent (α-actinin) images of neonatal rat cardiomyocytes (NRCMs) in the presence or absence of MMP9 inhibitor that were administered PBS or Ang II for 48 h. Bar=200 μm. (B) Statistical results for the cell surface area (CSA) of NRCMs. (C-E) The relative mRNA expression levels of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC) in NRCMs in the presence or absence of MMP9 inhibitor that were administered PBS or Ang II for 48 h. The intensities were normalized to the levels of GAPDH mRNA with the value in the PBS group defined as 1. * P<0.05 versus the control group, # P<0.05 versus the Ang II group. n=6 samples per experimental group

(E, F) Uev1A mediates negative feedback of NF-κB signaling pathway is dependent on TACE activity. Forty-eight hours after transfection, the cells were pre-treated with BB-2516 (20 μM) for 1 h, followed by TNFα (40 ng/ml) treatment for 2 h. Cell was fixed and stained with antibodies against HA and p65. The same cells were also stained with DAPI to reveal nuclei. White arrows indicate UEV1A-overexpressed cells. (F) Quantitative analysis of the experiments shown in (E). Percentage of cells displaying nuclear p65 is shown for UEV1A- overexpressed or vector-transfected cells. Each sample contains about 1000 cells. The significance of differences was assessed by two-tailed Student's test (***P < 0.001).

NOB modulates Cps1 mRNA and protein expression. a Total protein extracts were prepared from liver samples collected from the four diet/treatment groups of wild-type mice at the indicated circadian time points (n = 3). Western blotting analysis was performed using anti-CPS1 antibody. RC regular chow, HFD high-fat diet, Veh vehicle, NOB Nobiletin. The results shown are representative of three independent experiments. See Additional file 1: Figure S1A for quantitative analysis. b Immunohistochemical staining of CPS1 in liver sections from mice with the indicated diet and treatment at ZT2. c Real-time RT-PCR analysis of Cps1 in liver samples collected as in (a). Data are presented as mean ± SEM (n = 3). Two-way ANOVA with Bonferroni post-hoc tests shows significant statistical differences between HFD.Veh and other three groups (p < 0.0001). d Western blotting analysis of protein lysates of liver samples collected at ZT 6 and 18 from mice with the indicated diet and treatment (n = 3). HPD indicates high-protein diet. The images shown to the left are representative of three independent experiments. Quantitation of Western blots was carried out and the results, presented as mean ± SEM, are shown in the lower panel. Two-way ANOVA with Bonferroni post-hoc tests, RC vs. HPD, ***p < 0.001. e Real-time qPCR analysis was carried out using total RNAs extracted from the liver samples described in (d). The results are presented as mean ± SEM. Two-way ANOVA with Bonferroni post-hoc tests, RC vs. HPD, *p < 0.05.

(B) Florescent images of EpEX (green) and EpICD (red) in PK-15 cells, which were transfected by pSIN-fEpCAM-mCherry and treated with or without TAPI-1/DAPT. Nuclei were stained by Hoechst 33342 (blue). Scale bar, 25 μm.