Journal: OncoTargets and therapy

Article Title: miR-495 inhibits proliferation, migration, and invasion and induces apoptosis via inhibiting PBX3 in melanoma cells

doi: 10.2147/OTT.S152362

Figure Lengend Snippet: miR-495 inhibits melanoma cell proliferation, migration, invasion, and colony formation in vitro. ( A ) A375 and MeWo cells were transfected with miR-495 mimics, negative control (miR-NC), negative control inhibitor (NC inhibitor), or miR-495 inhibitor for 24 hours, then the expression of miR-495 was quantified by qRT-PCR. ( B ) miR-495 mimics, miR-NC, NC inhibitor or miR-495 inhibitor-transfected A375, and MeWo cells were subjected to MTT assay. ( C ) miR-495 mimics, miR-NC, NC inhibitor or miR-495 inhibitor-transfected A375, and MeWo cells were subjected to wound healing assay. ( D ) miR-495 mimics, miR-NC, NC inhibitor or miR-495 inhibitor-transfected A375, and MeWo cells were subjected to transwell assay. ( E ) miR-495 mimics, miR-NC, NC inhibitor or miR-495 inhibitor-transfected A375, and MeWo cells were subjected to colony formation assay. Data are presented as mean ± SD (n=3). All experiments were performed three times, and representative images are presented. * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: About 1×10 5 cells were plated on the upper Transwell chambers (Costar; Corning Incorporated, Corning, NY, USA) with Matrigel (BD Bioscience, San Jose, CA, USA).

Techniques: Migration, In Vitro, Transfection, Negative Control, Expressing, Quantitative RT-PCR, MTT Assay, Wound Healing Assay, Transwell Assay, Colony Assay

Journal: OncoTargets and therapy

Article Title: miR-495 inhibits proliferation, migration, and invasion and induces apoptosis via inhibiting PBX3 in melanoma cells

doi: 10.2147/OTT.S152362

Figure Lengend Snippet: Silencing of pre-B-cell leukemia transcription factor 3 (PBX3)-inhibited cell proliferation, migration, and invasion in melanoma cells. ( A ) A375 and MeWo cells were transfected with negative control siRNA (si-NC) or siRNA against PBX3 (si-PBX3) for 24 hours, the mRNA expression levels of PBX3 in both the cells were examined by RT-qPCR and ( B ) the protein levels of PBX3 in both the cells were examined by Western blot. ( C ) A375 and MeWo cells were transfected with si-NC or si-PBX3 for indicated time, and cell proliferation was analyzed by MTT assay. ( D ) A375 and MeWo cells were transfected with si-NC or by transwell assay. ( E ) Cell migration was analyzed by si-PBX3 for 24 hours, and cell invasion was analyzed by wound healing assay. Data are presented as mean ± SD (n=3). All experiments were performed three times, and representative images are presented. * P <0.05, ** P <0.01, *** P <0.001. Abbreviation: NC, negative control.

Article Snippet: About 1×10 5 cells were plated on the upper Transwell chambers (Costar; Corning Incorporated, Corning, NY, USA) with Matrigel (BD Bioscience, San Jose, CA, USA).

Techniques: Migration, Transfection, Negative Control, Expressing, Quantitative RT-PCR, Western Blot, MTT Assay, Transwell Assay, Wound Healing Assay

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Overexpressed MPS-1 contributes to endometrioma development through the NF-κB signaling pathway

doi: 10.1186/s12958-021-00796-z

Figure Lengend Snippet: MPS-1 regulates proliferation, migration, invasion, apoptosis and cell cycle of EcESCs. A Transfection efficiency of MPS-1 siRNA was detected by qRT-PCR. B A representative western blotting image of MPS-1 after EcESCs were transfected with MPS-1 siRNA for 72 h (n = 2). C Based on the results of western blotting, knockdown efficiency of MPS-1 siRNA was analyzed by Image J software. D The effects of MPS-1 siRNA on proliferation were evaluated by CCK-8 assay. E , G The representative images of EcESCs apoptosis and cell cycle were depicted by flow cytometry after MPS-1 siRNA transfection for 72 h. F , H The apoptosis analysis and cell cycle analysis were based on the results of flow cytometry. I , K Transwell assays were used to determine the effects of MPS-1 siRNA on migration and invasion. The scale bar is 50 μm. J , L Cell count analysis of Transwell migration and invasion. NC, si-MPS-1 EcESCs transfected with negative control siRNA or MPS-1 siRNA (n = 4). Data are presented as mean ± SEM (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001)

Article Snippet: After 48 h of transfection, 5 × 10 4 EcESCs in 200 μL DMEM/F12 were seeded onto the upper transwell chamber (BD Falcon, USA).

Techniques: Migration, Transfection, Quantitative RT-PCR, Western Blot, Knockdown, Software, CCK-8 Assay, Flow Cytometry, Cell Cycle Assay, Cell Counting, Negative Control

Journal: Microvascular research

Article Title: Semaphorin 5A promotes angiogenesis by increasing endothelial cell proliferation, migration, and decreasing apoptosis

doi: 10.1016/j.mvr.2009.10.005

Figure Lengend Snippet: Sema5A induces migration of endothelial cells through Met kinase activity. (A) Increased migration of HMEC-1 cells treated with Sema5A. Migration of HMEC-1 cells in response to Sema5A was examined using a transwell chamber assay (see Materials and methods for description). The values represent the average number of cells migrated±SEM. *Significantly different from HMEC-1 cells cultured in media alone (p<0.05). (B) HMEC-1 cells undergoing migration in response to Sema5A were captured using a light microscope at 200× magnification. HMEC-1 treated with a. media alone, b. 10 ng/ml of Sema5A and c. 10 ng/ml of VEGF-A. (C) Sema5A–induced migration of endothelial cells is mediated through Met receptor. Migration of HMEC-1 cells in response to Sema5A with or without neutralizing antibody for Met receptor was examined using a transwell chamber assay. The values are percent cells migrated in the presence of Sema5A with or without Met antibody ± SEM. *Significantly different from endothelial cells treated with Sema5A alone.

Article Snippet: HMEC-1 cells (1×10 5 ) were plated in duplicate onto the upper transwell chamber (6.5 mm, Corning Costar, Cambridge, MA).

Techniques: Migration, Activity Assay, Transwell Chamber Assay, Cell Culture, Light Microscopy

Journal: PLoS ONE

Article Title: 14-3-3β Promotes Migration and Invasion of Human Hepatocellular Carcinoma Cells by Modulating Expression of MMP2 and MMP9 through PI3K/Akt/NF-κB Pathway

doi: 10.1371/journal.pone.0146070

Figure Lengend Snippet: (A) Matrigel-free Transwell assay was conducted to assess the migration ability of SMCC-7721 and Hep3B cells that were transfected with 14-3-3β expression construct or the empty vector pcDNA3.1. (B) Matrigel-based Transwell assay was performed to examine the invasiveness of SMCC-7721 and Hep3B with exogenous expression of 14-3-3β. (C) Matrigel-free migration assay and (D) Matrigel-based invasion assay were performed in CSQT-2 cells with and without expression of 14-3-3β-specific siRNA. This figure shows the representative images and the statistical analysis of three independent experiments. Values are expressed as the mean ± standard deviation. *** p <0.001.

Article Snippet: Cell migration ability was assessed by the Transwell assay with uncoated upper chambers (Corning, New York, USA) [ ], whereas cell invasion was examined by the Transwell assay with the upper chamber pre-coated with phenol red-free Matrigel (BD Biosciences, San Jose, CA, USA).

Techniques: Transwell Assay, Migration, Transfection, Expressing, Construct, Plasmid Preparation, Invasion Assay, Standard Deviation

Journal: PLoS ONE

Article Title: 14-3-3β Promotes Migration and Invasion of Human Hepatocellular Carcinoma Cells by Modulating Expression of MMP2 and MMP9 through PI3K/Akt/NF-κB Pathway

doi: 10.1371/journal.pone.0146070

Figure Lengend Snippet: (A) Expression of 14-3-3β, MMP2 and MMP9 in 14-3-3β-transfected SMCC-7721 and Hep3B cells. (B) Western blot analysis of phosphorylation status of various signaling proteins and the level of nuclear p65 in 14-3-3β-overexpressing HCC cells. β-actin was used as the internal control for total or cytosolic proteins, while Lamin A served as the internal control for nuclear proteins. The 14-3-3β-transfected Hep3B cells were treated with (C) LY294002 (20 μM), the inhibitor of PI3K/Akt signaling, or (D) PDTC (10 μM), the inhibitor of NF-κB signaling, for 5 h, followed by western blot analysis of the expression of MMP2 and MMP9. (E) The invasiveness of 14-3-3β-overexpressing cells with and without LY294002 (20 μM) or PDTC (10 μM) treatment was assessed by Matrigel-based Transwell assay. Values are expressed as the mean ± standard deviation of three independent experiments. *** p <0.001. (F) Western blot analysis was performed to determine the activation status of p-Akt and NF-κB in 14-3-3β-overexpressing cells after treating the cells with LY294002 (20 μM) for 5 h.

Article Snippet: Cell migration ability was assessed by the Transwell assay with uncoated upper chambers (Corning, New York, USA) [ ], whereas cell invasion was examined by the Transwell assay with the upper chamber pre-coated with phenol red-free Matrigel (BD Biosciences, San Jose, CA, USA).

Techniques: Expressing, Transfection, Western Blot, Transwell Assay, Standard Deviation, Activation Assay