Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Avicularin Reduces the Expression of Mediators of Inflammation and Oxidative Stress in Bradykinin-Treated MG-63 Human Osteoblastic Osteosarcoma Cells

doi: 10.12659/MSM.921957

Figure Lengend Snippet: The effects of avicularin on the levels of MDA, SOD, and catalase in bradykinin-treated MG-63 human osteoblastic osteosarcoma cells. Increasing concentrations of avicularin treated MG-63 cells for 48 h, then we used 1 μM bradykinin to treat the cells for 24 h. ( A ) The malondialdehyde (MDA) content was determined by MDA assay kit. ( B ) Superoxide dismutase (SOD) activity was determined by the SOD assay kit. ( C ) Catalase activity was measured using a catalase assay kit. Data are presented as the mean±standard deviation (SD) from three independent experiments. ** p<0.01 vs. the control group; # , ## p<0.05, p<0.01 vs. the bradykinin group.

Article Snippet: Superoxide dismutase (SOD) activity in the MG-63 was measured using the SOD assay kit (Cayman Chemical Company, Ann Arbor, MI, USA), according to the manufacturer’s instructions.

Techniques: Multiple Displacement Amplification, Activity Assay, Standard Deviation, Control

Journal: eLife

Article Title: FXR1 regulates transcription and is required for growth of human cancer cells with TP53/FXR2 homozygous deletion

doi: 10.7554/eLife.26129

Figure Lengend Snippet: ( A ) Copy number alterations in TP53, FXR2, FXR1, and FMR1 in various types of human tumors based on TCGA data analysis. ( B ) Schematic diagram of FXR2 and TP53 location at chromosome 17p13.1. ( C ) Cell proliferation rate upon FXR1 inducible knockdown. Measurements of change in cell proliferation rate induced by control shRNA (shCtrl) or by FXR1 shRNA (FXR1-sh2, FXR1-sh3) upon doxycycline (Dox) treatment (at indicated days) in TP53 and FXR2 co-deletion-containing cancer cell lines HL-60 and H358 (upper panel) and in the copy-number-normal cancer cell lines MKN45 and AGS (lower panel) using an MTS assay. Data represent the mean ± s.d. of three independent experiments. ( D ) Rescuing FXR1-sh3-induced anti-proliferation by ectopic expression of FXR1 or FXR2 in TP53 / FXR2 deletion cancer cell line KATOIII. Upper panel: protein levels of a shRNA-resistant form of full-length FXR1 (FXR1m_a) or of FXR2 upon knockdown of endogenous FXR1 in a Western Blot (WB) assay. Lower panel, cell proliferation in vector, FXR1m_a, or FXR2 ectopic expression cells upon Dox-induced FXR1 knockdown in the MTS assay. Data represent the mean ± s.d. of three independent experiments. ( E ) Rescuing FXR1-sh3-induced anti-proliferation by ectopic expression of FMR1 in the TP53 / FXR2 deletion cancer cell line H358. Upper panel: ectopic expression level of FMR1 and knockdown efficiency of FXR1 by Dox-induced shRNA. Lower panel, cell proliferation in H358 cells ectopically expressing vector or FMR1 upon Dox-induced FXR1 knockdown in the MTS assay. Data represent the mean ± s.d. of three independent experiments. ( F ) Cell proliferation change upon FXR1-inducible knockdown in cancer cells with a TP53 single deletion (H1299, L540, MG-63, SKOV3) or an FXR2 single deletion (Hep3B). Upper panel, FXR1 protein level upon shCtrl or FXR1-sh3 treatment. Lower panel, cell proliferation measurement in MTS assay. Data represent the mean ± s.d. of three independent experiments. ( G ) Tumor growth in cancer cell xenograft upon FXR1 knockdown. Left, growth curves of xenograft tumors derived from subcutaneously implanted HL-60 cells stably expressing shCtrl or FXR1-sh3 upon Dox treatment. Tumor volume (mm ) represents the mean ± s.e.m of six mice for each group after the indicated number of days with Dox treatment. Right, the tumor sizes on the termination day. The cell proliferation rate was determined by measuring absorbance at 490 nm in the MTS assay (Y axis). *p<0.05, **p<0.01, ***p<0.001. Also see , , , , and . 10.7554/eLife.26129.010 Figure 1—source data 1. Source data for .

Article Snippet: KATOIII, HL-60, H358, MKN45, AGS, HepG2, A549, H1299, MG-63, SKOV3, Hep3B, and HEK 293 T cell lines were purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA, http://www.atcc.org ).

Techniques: Knockdown, Control, shRNA, MTS Assay, Expressing, Western Blot, Plasmid Preparation, Derivative Assay, Stable Transfection

Journal: eLife

Article Title: FXR1 regulates transcription and is required for growth of human cancer cells with TP53/FXR2 homozygous deletion

doi: 10.7554/eLife.26129

Figure Lengend Snippet: Summary of FXR1-knockdown-induced anti-proliferation in cancer cell lines and CRISPR-Cas9-engineered cell clones. The cell line or CRISPR clone name, cancer type, TP53 and FXR2 copy number alteration (based on the CCLE database or knockout confirmation), FXR1-sh3 knockdown (KD) efficiency (determined by q-RT-PCR and WB), and FXR1 KD-induced anti-proliferation efficiency are listed. Dark green, anti-proliferation efficiency >60%. Yellow green, anti-proliferation 30–60%. Yellow, anti-proliferation <30%. Data of KD efficiency and anti-proliferation efficiency represent means of at least three independent experiments. Del, deletion; WT, copy number normal; KO, knockout. 10.7554/eLife.26129.016 Table 1—source data 1. Source data for Table 1 .

Article Snippet: KATOIII, HL-60, H358, MKN45, AGS, HepG2, A549, H1299, MG-63, SKOV3, Hep3B, and HEK 293 T cell lines were purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA, http://www.atcc.org ).

Techniques: CRISPR, Clone Assay, Knock-Out, Knockdown