Journal: British journal of cancer

Article Title: Inactivation of the WASF3 gene in prostate cancer cells leads to suppression of tumorigenicity and metastases.

doi: 10.1038/sj.bjc.6605850

Figure Lengend Snippet: Figure 7 Knockdown of WASF3 expression suppresses MMP9 expression levels and inhibits MMP9 release from prostate cancer cell lines. (A) RT–PCR analysis of PC3 and DU145 cells showing knockdown of WASF3 also show down-regulation of MMP9, but not MMP2 expression levels. Cont ¼ parental cells. (B) Secretion of the pro- and active form of MMP9 from PC3 and DU145 cells were measured in the supernatants of actively growing cells using the Fluorokine MAP assay procedure. Cells from each clone showing knockdown of WASF3 also showed reduced MMP9 levels compared with parental (control) and non-knockdown cells that did not.

Article Snippet: After 24 h, the media were collected in tubes and centrifuged for 10 min at 10 000 g. The pro- and active MMP-9 levels released into the media were measured using a Fluorokine MAP human MMP9 kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Knockdown, Expressing, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Scientific Reports

Article Title: All trans retinoic acid alleviates coronary stenosis by regulating smooth muscle cell function in a mouse model of Kawasaki disease

doi: 10.1038/s41598-021-93459-3

Figure Lengend Snippet: Immunohistochemical staining of α-SMA (upper), proliferating cell nuclear antigen (PCNA) (middle), and matrix metalloproteinase-9 (MMP-9) (bottom) in mice with PBS, LCWE and LCWE + atRA (a) . All positive cells are shown as a brown color. Isotype antibodies were used as negative control; α-SMA: mouse polyclonal IgG2a kappa, PCNA: rabbit IgG, MMP-9: mouse monoclonal IgG1a kappa. Scale bar = 100 μm. The number of intimal αSMA- (b) and PCNA-positive cells (c) and the intimal MMP-9-positive area (d) in mice treated with PBS (n = 5), LCWE (n = 6) and LCWE + atRA (n = 7) were quantitatively analyzed and compared among the groups. The data are expressed as the mean ± SD. *p < 0.0001. All the data were generated from 1 experiment at the same timeframe.

Article Snippet: The cells were then incubated with medium alone, PDGF-BB (20 ng/ml) without atRA or in the presence of atRA at 10 nM for the next 24 h. Cell culture supernatants were collected and total MMP-9 activity (MMP-9 activity assay kit for humans, Cosmo Bio Co., Ltd, Tokyo, Japan) and TNF-α concentration (Human TNF-α immunoassay, R&D Systems, Inc. MN, USA) were measured according to the manufacturer's instructions.

Techniques: Immunohistochemical staining, Staining, Negative Control, Generated

Journal: Scientific Reports

Article Title: All trans retinoic acid alleviates coronary stenosis by regulating smooth muscle cell function in a mouse model of Kawasaki disease

doi: 10.1038/s41598-021-93459-3

Figure Lengend Snippet: Effect of atRA on LCWE-induced elastin breaks. EVG staining of CAs from mice treated with PBS, LCWE and LCWE + atRA (a) . Elastin breaks in the external elastic lumina (EEL) (b) and in the internal elastic lumina (IEL) (c) among the groups (PBS: n = 5, LCWE: n = 6, LCWE + atRA: n = 7). (d) The serum level of MMP-9 in mice with PBS (n = 5), LCWE (n = 6) and LCWE + atRA (n = 7). The data are expressed as the mean ± SD. *p < 0.0001, **p < 0.05. All the data were generated from 1 experiment at the same timeframe.

Article Snippet: The cells were then incubated with medium alone, PDGF-BB (20 ng/ml) without atRA or in the presence of atRA at 10 nM for the next 24 h. Cell culture supernatants were collected and total MMP-9 activity (MMP-9 activity assay kit for humans, Cosmo Bio Co., Ltd, Tokyo, Japan) and TNF-α concentration (Human TNF-α immunoassay, R&D Systems, Inc. MN, USA) were measured according to the manufacturer's instructions.

Techniques: Staining, Generated

Journal: Scientific Reports

Article Title: All trans retinoic acid alleviates coronary stenosis by regulating smooth muscle cell function in a mouse model of Kawasaki disease

doi: 10.1038/s41598-021-93459-3

Figure Lengend Snippet: The migration of human coronary artery smooth muscle cells (HCASMCs). A total of 10,000 cells were incubated with PDGF-BB (20 ng/ml) to induce cell migration in the presence or absence of various concentrations of atRA (0.1, 1, and 10 nM) for 72 h (a) . Dotted circles indicate covered areas where the cells were not attached at baseline. The cells that migrated from the uncovered area (μm 2 ) to the covered area were quantitatively measured in each group (b) . Magnification × 4. *p < 0.05 vs Medium, ǂp < 0.0001 vs PDGF-BB, atRA10 nM, †p < 0.01 vs PDGF-BB, Medium. HCASMCs (1 × 10 6 cells) were placed in 60 mm culture dishes and incubated with PDGF-BB (20 ng/ml) in the presence or absence of 10 nM atRA for 24 h. Total MMP-9 activity and TNF-α concentration were measured in each group (c) . *p < 0.05. The values are expressed as the mean ± SD of quadruplicate determinations in 2 independent experiments.

Article Snippet: The cells were then incubated with medium alone, PDGF-BB (20 ng/ml) without atRA or in the presence of atRA at 10 nM for the next 24 h. Cell culture supernatants were collected and total MMP-9 activity (MMP-9 activity assay kit for humans, Cosmo Bio Co., Ltd, Tokyo, Japan) and TNF-α concentration (Human TNF-α immunoassay, R&D Systems, Inc. MN, USA) were measured according to the manufacturer's instructions.

Techniques: Migration, Incubation, Activity Assay, Concentration Assay