Multivariate analysis with stepwise linear regression of factors independently associated with interstitial and glomerular fibrosis.

<p>Variables included p less than 0.1 in univariate linear regression analysis.</p><p><i>p</i><0.05 was considered statistically significant.</p

Expression of MMP-9 in different renal tissues between patients with and without cancer.

<p>Abbreviation: Gc, glomerular cytoplasm; NTn, normal tubular nucleus; NTc, normal tubular cytoplasm; ATn, atrophic tubular nuclei; ATc, atrophic tubular cytoplasm.</p><p>Data were analyzed by the chi-squared test and Fisher’s exact test accordingly and <i>p</i><0.05 indicates significance.</p

Representative panels showing different expression intensity of MMP-9 in atrophic tubular nuclear compared to normal tubular cytoplasm nearby fibrotic renal parenchyma.

<p>(A) no fibrosis with increased cytoplasm stain in normal tubules, (B) mild fibrosis with decreased cytoplasm stain in normal tubules and increased nuclear stain in atrophy tubules, (C) severe fibrosis with decreased cytoplasm stain in normal tubules, and (D) severe fibrosis with increased nuclear stain in atrophy tubules. (IHC stain, x 20).</p

Associations between interstitial and glomerular fibrosis with clinicopathologic variables.

<p>Interstitial fibrosis scores were significantly correlated with glomerular fibrosis scores (r = 0.741, <i>p</i><0.001).</p><p><i>p</i><0.05 was considered statistically significant.</p

Intensity of MMP-9 expression in different regions of renal tissues divided by low and high interstitial fibrosis score (left) and glomerular fibrosis score (right).

<p>NTn, normal tubular nucleus; NTc, normal tubular cytoplasm; Gn, glomerular nuclei; Gc, glomerular cytoplasm; ATn, atrophic tubular nuclei; ATc, atrophic tubular cytoplasm.</p><p>Data were analyzed by the chi-squared test and <i>p</i><0.05 indicates significance.</p

Demographic and clinical characteristics of patients divided by low and high renal interstitial fibrosis score (left) and glomerular fibrosis score (right).

<p>BMI, body mass index; CKD, chronic kidney disease; DM, diabetes mellitus; eGFR, estimated glomerular filtration; HTN, hypertension; RCC, renal cell carcinoma; UCC, urothelial cell carcinoma, TCH, total cholesterol.</p><p><i>p</i><0.05 indicates significance.</p

Associations between intensity of MMP-9 expression and cancer over different renal tissues.

<p><i>p</i><0.05 was considered statistically significant.</p

Effect of LicA on the MAPK pathway in HCC cells.

<p>HCC cells were treated with various concentrations (0, 5, 10, and 20 µM) of LicA for 24 h, and then cell lysates were subjected to Western blotting with anti-p-ERK, anti-ERK, anti-p-p38, anti-p-38, anti-p-JNK, anti-JNK, and β-actin antibodies. β-actin was used as the loading control. Data are presented as the mean±SE of at least three independent experiments. <i>*p</i><0.05, compared with that of the untreated control (0 µM).</p

Effect of LicA on the viability, migration, and invasion of HCC cells.

<p>(A) SK-Hep-1, (B) HA22T/VGH and (C) normal hepatic cells (4×10⁴ cells/mL) were treated with various concentrations (0, 5, 10, and 20 µM) of LicA for 24 and 48 h. Cell viability was determined by using an MTT assay. (D) For the Boyden chamber migration assay, cells were treated with various concentrations of LicA for 24 h, and then cell migration was measured using a Boyden chamber for 12 h with polycarbonate filters (8 µm). (E) Cell invasion was measured using the Boyden chamber for 24 h; the polycarbonate filters were pre-coated with Matrigel. Motility was quantified by counting the number of cells that migrated to the undersides of the membrane under microscopy (200×). Data are presented as the mean±SE of at least three independent experiments. *<i>p</i><0.05, compared with that of the untreated control (0 µM).</p

Effect of MKK4 on uPA expression levels, as well as the migration and invasion SK-Hep-1 cells.

<p>(A) Cells were treated with various concentrations (0, 5, 10, and 20 µM) of LicA for 24 h, and then cell lysates were subjected to SDS-PAGE followed by Western blotting with anti-p-MKK3/6, anti-MKK3/6, anti-p-MKK4, and anti-MKK4 antibodies. β-actin was used as an internal control. (B) Cells were treated with si-MKK4 (200 nM), and its inhibitory potency toward MKK4 or MKK3/6 was analyzed with Western blotting. (C) SK-Hep-1 cells were pretreated with si-MKK4 (200 nM) for 24 h and incubated in the presence or absence of LicA (10 µM) for another 24 h. Cell lysates were then subjected to western blotting to determine the protein expression levels of uPA. Cells were also assessed for migration (D) and invasion (E). *<i>p</i><0.05, untreated cells versus si-MKK4 or LicA; # <i>p</i><0.01, LicA versus si-MKK4 plus LicA.</p