Journal: Experimental and Therapeutic Medicine

Article Title: Effects of CoCl 2 -simulated hypoxia on the expression levels of matrix metalloproteinases in renal adenocarcinoma cells and renal tubular epithelial cells

doi: 10.3892/etm.2018.6337

Figure Lengend Snippet: (A-F) Effects of chemical hypoxia on the expression levels of MMPs differed. (A, C and E) Expression levels of RECK, MMP2, MMP9 and MMP14 in HK2, 786-0 and HMEC-1 cells were determined by western blot analysis after incubation with varying concentrations of CoCl2 from 0 to 200 µmol/l for 24 h. (B, D and F) Normalized densitometric quantitation. *P<0.05, #P>0.05 as indicated. In all cases, at least three independent experiments were performed. MMP, matrix metalloproteinase; HK2, human renal tubular epithelial cells; 786-0, human renal cell cancer cells; HMEC-1, human microvascular endothelial cells.

Article Snippet: The source and dilution of unconjugated antibodies used in this study are as follows: HIF-1α (rabbit pAb; NB100-134SS, 1:1,000; Novus Biologicals, Ltd., Cambridge, UK), RECK (goat pAb; AF1734, 1:1,000; R&D Systems, Inc., Minneapolis, MN, USA), MMP2 (rabbit pAb; NB200-193, 1:1,000; Novus Biologicals), MMP9 (rabbit mAb; ab76003, 1:1,000; Abcam, Cambridge, UK), MMP14 (rabbit mAb; ab51074, 1:1,000; Abcam), TIMP2 (rabbit pAb; BS1366, 1:500; Bioworld Technology, Inc., St. Louis Park, MN, USA), VEGF-A (rabbit pAb; BS2431, 1:500; Bioworld Technology, Inc.), β-Tubulin (Mouse pAb; BS1482 M, 1:1,000; Bioworld Technology, Inc.), Fluorescein (FITC)-conjugated Goat Anti-Rabbit IgG (BS10950, 1:200; Bioworld Technology, Inc.), Horseradish Peroxidase (HRP)-conjugated Goat Anti-Rabbit IgG (BS13278, 1:3,000; Bioworld Technology, Inc.), Horseradish Peroxidase (HRP)-conjugated Goat Anti-Mouse IgG (BS12478, 1:3,000; Bioworld Technology, Inc.), Horseradish Peroxidase (HRP)-conjugated Rabbit Anti-Goat IgG (BS30503, 1:3,000; Bioworld Technology, Inc.).

Techniques: Expressing, Western Blot, Incubation, Quantitation Assay

Journal: OncoTargets and Therapy

Article Title: miR-590-5p regulates gastric cancer cell growth and chemosensitivity through RECK and the AKT/ERK pathway

doi: 10.2147/ott.s110923

Figure Lengend Snippet: Figure 6 Reintroduction of RECK inhibited the effect of miR-590-5p on SGC-7901 cells. Notes: (A) After transfection of RECK (lacking 3′-UTR), the protein level of RECK was restored. MTT (B) and invasion assays (C) showed the inhibitory effects of RECK on GC cells. (D) RECK increased the sensitivity of GC cells to cDDP and PTX. (E) The p-AKT, p-ERK, and pSTAT3 expression was decreased by restoration of RECK. *P,0.05; **P,0.01; ***P,0.001. Abbreviations: cDDP, cisplatin; OD, optical density; GC, gastric cancer; 3′-UTR, 3′ untranslated region; PTX, paclitaxel; Mim con, mimics control.

Article Snippet: The membranes were blocked by incubation in Tris-buffered saline with Tween 20 (25 mM Tris, pH 7.4, 150 mM NaCl, 0.1% Tween 20) containing 5% fat free milk for 1 hour at room temperature and were then immunoblotted with primary antibodies against RECK (R&D Systems, Inc., Minneapolis, MN, USA), AKT, p-AKT (Ser473), ERK, p-ERK (Cell Signaling Technology, Danvers, MA, USA), pSTAT3, STAT3, and β-actin (Affinity, Ossipee, NH, USA), followed by incubation with HRP-conjugated secondary antibodies (Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: Transfection, Expressing, Control