Fig 2: JNK, p38, and ERK inhibitors reduced H. pylori-induced MMP-10 expression in AGS cells. The cells were pretreated with SB600125 (a JNK inhibitor, 20 µM) or SB203580 (a p38 inhibitor, 20 µM) for 60 min, or U0126 (an ERK inhibitor, 10 µM) for 30 min, and then infected with H. pylori for 24 h. MMP-10 levels were determined by Western blot analysis. Actin was used as a loading control.

Fig 3: ß-Carotene inhibits H. pylori-induced activation of MAPKs and AP-1, and expression of MMP-10 in AGS cells. Cells were pretreated with the indicated concentrations of ß-carotene for 2 h and then infected with H. pylori for 24 h (A,B), 1 h (C, left panel), 30 min (C, right panel), and 1 h (D). (A) MMP-10 mRNA expression was analyzed by real-time PCR and normalized to ß-actin mRNA. All data are shown as the mean ± S.E. of three independent experiments. * p < 0.05 vs. none (cells without any treatment or infection); + p < 0.05 vs. control (cells with H. pylori infection alone). (B) The level of MMP-10 was determined by Western blot analysis, using actin as the loading control. (C) Protein levels of phosphorylated or total form of JNK1/2, p38, and ERK1/2 were determined by Western blot analysis. Actin served as a loading control (upper panel). Lower panel: The densitometry data represent means ± S.E. from three immunoblots and are shown as relative density of phosphorylated protein band normalized to total form of protein level. * p < 0.05 vs. none (cells without any treatment or infection); + p < 0.05 vs. control (cells with H. pylori infection alone). (D) The DNA binding activity of AP-1 was measured by an electrophoretic mobility shift assay (EMSA).

Fig 4: MMP10 is required in the ghrelin-Aurora A signaling axis to promote ccRCC invasion. (A) Kaplan–Meier plot showing the association of indicated MMP with RCC (TCGA) patient survival was represented. Data were retrieved from Human Protein Atlas website. (B) Representative Aurora A and MMP10 expression pattern in ACHN cells overexpressing ghrelin combined with Aurora A or MMP10 knockdown respectively. (C) Alternations in relative ACHN cell invasion ability were shown. Magnification: 100× (D) The regulation of ghrelin toward MMP10 expression was further studied in ACHN cells silencing GHS-R1a. Figures were represented from the results of three repeated experiments with similar pattern. ** p < 0.01, *** p < 0.001.