Journal: The Journal of Clinical Investigation

Article Title: Wilms tumor 1 impairs apoptotic clearance of fibroblasts in distal fibrotic lung lesions

doi: 10.1172/JCI188819

Figure Lengend Snippet: ( A ) Schematic representation of the animal experiment involving PDGFR α CreERT (control) and PDGFR α CreERT WT1 OE (cWT1 OE ) mice. Mice were treated repeatedly with bleomycin via intratracheal administration and with tamoxifen via intraperitoneal injection. ( B ) Quantification of Wt1 gene transcripts by RT-PCR in total lung RNA isolated from control and cWT1 OE mice ( n = 6/group). ** P < 0.01, by 2-tailed Student’s t test. ( C ) Representative confocal images of lung sections from control and cWT1 OE mice co-immunostained for WT1 (red), vimentin (green), and DAPI (blue). Scale bars: 20 μm. ( D ) Representative images of Masson’s trichrome–stained lung sections from control and cWT1 OE mice. Original magnification, ×4 and ×20, respectively. Scale bars: 1,500 μm and 200 μm, respectively. ( E ) The percentage of fibrotic area was quantified in control and cWT1 OE mice using BZ-X image analysis ( n = 6/group). **** P < 0.0001, by 2-tailed Student’s t test. ( F ) Hydroxyproline levels were measured in the right lungs of control and cWT1 OE mice ( n = 6/group). *** P < 0.001, by 2-tailed Student’s t test. ( G ) Lung resistance was assessed using FlexiVent in control and cWT1 OE mice treated with bleomycin ( n = 6/group). * P < 0.01, by 2-tailed Student’s t test. ( H ) Proliferation of fibroblasts was quantified using a BrdU incorporation assay in lung cultures from control and cWT1 OE mice treated with bleomycin ( n = 3/group). * P < 0.05, by 2-tailed Student’s t test. ( I ) Fibroblasts from the lung cultures of control and cWT1 OE mice treated with bleomycin were treated with anti-Fas antibody for 24 hours, followed by TUNEL staining (red). Representative confocal images are shown; nuclei are stained with DAPI (blue). Original magnification, ×20. Scale bars: 100 μm. ( J ) Percentage of TUNEL + fibroblasts in total DAPI + fibroblasts ( n = 3/group). * P < 0.05 and ** P < 0.01, by 1-way ANOVA. Data are representative of 2 independent experiments with similar findings.

Article Snippet: Cell proliferation was evaluated using the BrdU kit (Cell Signaling Technology) as described previously ( ).

Techniques: Control, Injection, Reverse Transcription Polymerase Chain Reaction, Isolation, Staining, BrdU Incorporation Assay, TUNEL Assay

Journal: PLoS ONE

Article Title: Alpha-Catulin Contributes to Drug-Resistance of Melanoma by Activating NF-κB and AP-1

doi: 10.1371/journal.pone.0119402

Figure Lengend Snippet: ( A ) Stable infected Mel.7 cells (n.s., sh-catu2) were treated with 0, 5, 10 and 20 μg/ml cisplatin for 24 h and analyzed by Western blot with antibody against Ki67. α-Tubulin was used as a loading control and quantification was performed with BioRad software. ( B ) Mel.7 cells (n.s., sh-catu2) were treated with 20, 10, 5, or 0 μg/ml cisplatin for 48 h and BrdU assay was performed. Therefore, cells were stained with BrdU solution and antibodies against BrdU and HRP conjugated secondary antibody was detected at 450 nm using a multiplate reader. ( C ) Mel.7 cells (n.s., sh-catu2) were treated with 0 or 10 μg/ml cisplatin for 18 hours, fixed, stained with propidium-iodide solution and analysed for cell cycle distribution using flow cytometry. ( D ) Cells as in ( C ) were analysed using western blot with antibodies against p21 cip/waf and p53.

Article Snippet: After 24 hours cells were treated with 20, 10, or 5 μg/ml cisplatin or were left untreated and after 48 hours CytoSelect BrdU Cell Proliferation ELISA Kit (Cell Biolabs, Inc.) was used to determine cell proliferation.

Techniques: Infection, Western Blot, Control, Software, BrdU Staining, Staining, Flow Cytometry

Journal: Scientific Reports

Article Title: Lamprey Prohibitin2 Arrest G2/M Phase Transition of HeLa Cells through Down-regulating Expression and Phosphorylation Level of Cell Cycle Proteins

doi: 10.1038/s41598-018-22212-0

Figure Lengend Snippet: Effect of Lm-PHB2 on the cell cycle. The double staining with both APC-BrdU and PI for detection of G1/S and/or G2/M cell cycle alteration after HeLa cells treated with rLm-PHB2 protein and PBS treatment as control.

Article Snippet: Cell cycle detection kit and BrdU cell proliferation detection kit was purchased from KeyGEN BioTECH.

Techniques: Double Staining, Control

Journal: Oncotarget

Article Title: Long-term prognostic implications and therapeutic target role of hexokinase II in patients with nasopharyngeal carcinoma

doi: 10.18632/oncotarget.7116

Figure Lengend Snippet: Cells without any treatments were adopted as controls. ( A ) 3-BrOP reduces intracellular ATP concentration of NPC cells. ATP levels were determined using the ATPlite kit according to manufacturer's protocol (PerkinElmer, Boston, MA). ( B ) Growth inhibitory effect of 3-BrOP on NPC cells. The cell proliferation was detected by BrdU in different cell groups.

Article Snippet: BrdU assay was performed using the BrdU Cell Proliferation assay kit (Merck, Marmstadt, Germany) following the manufacturer's protocol.

Techniques: Concentration Assay