Journal: The Journal of Biological Chemistry

Article Title: A potential strategy for reducing cysts in autosomal dominant polycystic kidney disease with a CFTR corrector

doi: 10.1074/jbc.RA118.001846

Figure Lengend Snippet: Cell proliferation is reduced. A and B, proliferation in kidneys of Pkd1fl/fl;Pax8rtTA;TetO-cre mice. Representative images of Ki67 (a cellular marker for proliferation) staining of PN21 kidney sections from DMSO- and VX-809–treated mice. Arrows indicate Ki67-positive cells. Pictures were acquired with a Zeiss microscope equipped ×20 objective. C, summary data for Ki67-positive cells. Columns represent averages ± S.E. of DMSO (vehicle)-treated (n = 3) and VX-809–treated (n = 3) mice. Statistical analysis was performed using a two-tailed Student's t test. D, proliferation in PN cells. PN cells were treated with VX-809 or DMSO. The bromodeoxyuridine (BrdU) concentration in the cells was measured by using a BrdU cell proliferation assay kit (MilliporeSigma 2750) according to the manufacturer's protocol. Columns represent averages ± S.E. for the OD of BrdU at 450/550 nm. Data were analyzed using Student's t test with n = 5–7. Methods were described previously (25). *, p > 0.05;**, p < 0.01.

Article Snippet: The bromodeoxyuridine (BrdU) concentration in the cells was measured by using a BrdU cell proliferation assay kit (MilliporeSigma 2750) according to the manufacturer's protocol.

Techniques: Marker, Staining, Microscopy, Two Tailed Test, Concentration Assay, BrdU Cell Proliferation Assay

Journal: Journal of Biomedical Science

Article Title: Intermittent hypoxia-induced protein phosphatase 2A activation reduces PC12 cell proliferation and differentiation

doi: 10.1186/1423-0127-21-46

Figure Lengend Snippet: Intermittent hypoxia (IH) effects on PC12 cell numbers, cell viability, cell proliferation and cell cycle progression. (A) Numbers of PC12 cells as evaluated using Hoechst staining (blue) and confocal microscopy. (B) Quantitative PC12 cell numbers after exposure to normoxia (RA) and IH for 1–4 days (n = 5 for per group). (C) PC12 cell numbers as determined by MTT assay after exposure to RA and IH for 1–4 days (n = 5 for per group). (D) PC12 cell proliferation determined by BrdU cell proliferation ELISA assay kit after exposure to 4 days of RA (RA4, n = 8) and IH (IH4, n = 8). (E) PC12 cell cycle progression after exposure to RA4 (n = 8), IH3 (n = 7) and IH4 (n = 7) as evaluated by propidium iodide staining and flow cytometry. Percentages of cells in G 0 /G 1 phase arrest (E and F). * p < 0.05 compared with RA in (B) and (C) or RA4 in (D) and (F) . Values are means ± SEMs.

Article Snippet: Cell proliferation was assessed using a BrdU cell proliferation ELSIA assay kit (cat. no. 2750, Millipore, USA).

Techniques: Staining, Confocal Microscopy, MTT Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Journal of Biomedical Science

Article Title: Intermittent hypoxia-induced protein phosphatase 2A activation reduces PC12 cell proliferation and differentiation

doi: 10.1186/1423-0127-21-46

Figure Lengend Snippet: Effect of intermittent hypoxia (IH)-induced ROS generation on PP2A activation and downregulated ERK1/2 activation leading to PC12 cell proliferation inhibition. (A) PC12 cell numbers determined by MTT assay after exposure to normoxia for 4 days (RA4, control, n = 16), and RA4 along with the ERK1/2 phosphorylation inhibitors U0126 (n = 8) and PD98059 (n = 7), the ERK1/2 phosphorylation activator nicotine (n = 14), and exposed to IH for 4 days (IH4, control, n = 8), IH4 along with superoxide dismutase (SOD, n = 12), the PP2A activation inhibitors 1,10-phenanthroline (Phe, n = 12), okadaic acid (OKA, n = 9) and cantharidin (Can, n = 9) and nicotine (n = 9). (B) PC12 cell proliferation determined by BrdU cell proliferation ELISA assay kit after exposure to RA4 (n = 8), IH4 (control, n = 8) and IH4 along with SOD (n = 7), Phe (n = 7), OKA (n = 7), Can (n = 7) and nicotine (n = 6). (C) Percentages of PC12 cells in G 0 /G 1 phase arrest as determined by propidium iodide staining and flow cytometry after exposure to RA4 (control, n = 8), RA4 along with U0126 (n = 4), RA4 with nicotine (n = 4), IH4 (control, n = 7), IH4 along with SOD (n = 6), Phe (n = 5), OKA (n = 5) and nicotine (n = 4). * p < 0.05 compared with RA4. # p < 0.05 compared with IH4. Values are means ± SEMs.

Article Snippet: Cell proliferation was assessed using a BrdU cell proliferation ELSIA assay kit (cat. no. 2750, Millipore, USA).

Techniques: Activation Assay, Inhibition, MTT Assay, Control, Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: CHIP functions as an oncogene by promoting colorectal cancer metastasis via activation of MAPK and AKT signaling and suppression of E-cadherin

doi: 10.1186/s12967-018-1540-5

Figure Lengend Snippet: CHIP silencing inhibits cell growth of DLD-1 cells. a The cell growth curves of the siCHIP and sictrl cells were detected using a real-time x-Celligence system. E-plate was plated with 10,000 cells/well and the cell growth was continuous monitored for 72 h. Significant differences were indicated (Student’s t test, ** P < 0.01, *** P < 0.001). b The apoptosis of the siCHIP and sictrl cells was determined by the AnnexinV/PI staining. AnnexinV positive and AnnexinV/PI positive cells were determined as apoptosis cells. Values ± SD from three individual experiments were displayed. c Cell proliferation of the siCHIP and sictrl cells was determined by the Brdu proliferation assay. 96-well plate was plated with 10,000 cells/well and was added 10 μl buffer after cultured for 24, 48 and 72 h. OD450 was measured using spectrophotometer microplate reader. Significant differences were indicated (Student’s t test,*** P < 0.001). d CHIP depletion caused cell cycle arrest at the G0–G1 phase. Cell cycle parameters of siCHIP and sictrl cells were determined by flow cytometry analysis using propidium iodide. The histograms report the percentage of cells in G0/G1, S and G2/M phases of all the establishing cell lines. *** P < 0.001. e Western blotting analysis of the protein expression of total ERK1/2, p-ERK1/2, p38, p-p38, as well as total AKT andp-AKT 473 and p-AKT 308 in siCHIP and sictrl cells. Actin was used as an internal control. f Western blotting analysis of the protein expression of the NF-κB subunits. Actin was used as an internal control. g Western blotting analysis of the protein expression of the cell cycle-related protein cyclinD1. Actin was used as an internal control

Article Snippet: The BrdU Cell Proliferation Assay Kit (Cat Nr. 2750, Merck Millipore, German) was used to detect the proliferation of CRC cells according to the manufacture’s instruction.

Techniques: Staining, Proliferation Assay, Cell Culture, Spectrophotometry, Flow Cytometry, Western Blot, Expressing, Control

Journal: Journal of Translational Medicine

Article Title: CHIP functions as an oncogene by promoting colorectal cancer metastasis via activation of MAPK and AKT signaling and suppression of E-cadherin

doi: 10.1186/s12967-018-1540-5

Figure Lengend Snippet: CHIP overexpression did not affect cell growth of DLD-1 cells. a The cell growth curves of the hCHIP and ctrl cells were detected using a real-time x-Celligence system. E-plate was plated with 10,000 cells/well and the cell growth was continuous monitored for 72 h. b The apoptosis of the hCHIP and ctrl cells was determined by the AnnexinV/PI staining. AnnexinV positive and AnnexinV/PI positive cells were determined as apoptosis cells. Values ± SD from three individual experiments were displayed. c Cell proliferation of the hCHIP and ctrl cells were determined by the Brdu proliferation assay. 96-well plate was plated with 10,000 cells/well and was added 10 μl buffer after cultured for 24, 48 and 72 h. OD450 was measured by means of spectrophotometer microplate reader. d Cell cycle parameters of hCHIP and ctrl cells were determined by flow cytometry analysis using propidium iodide. The histograms report the percentage of cells in G0/G1, S and G2/M phases of all the establishing cell lines. e Western blot analysis of the protein expression of total ERK1/2, p-ERK1/2, p38, p-p38 as well as total AKT and p-AKT 473 and p-AKT 308 in hCHIP and ctrl cells. Actin was used as an internal control. f Western blot analysis of the protein expression of the NF-κB subunits. Actin was used as an internal control. g Western blot analysis of the protein expression of the cell cycle related proteins cyclinD1. Actin was used as an internal control

Article Snippet: The BrdU Cell Proliferation Assay Kit (Cat Nr. 2750, Merck Millipore, German) was used to detect the proliferation of CRC cells according to the manufacture’s instruction.

Techniques: Over Expression, Staining, Proliferation Assay, Cell Culture, Spectrophotometry, Flow Cytometry, Western Blot, Expressing, Control