Lb 100

The IOMM-Lee human meningioma cell line was purchased from the American Type Culture Collection (ATCC, Rockville, MD, United States) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Biosharp, China) supplemented with 10% fetal bovine serum (Gibco, Auckland, United States) in a humidified incubator at 37°C with 5% CO₂.
LB-100 was provided by Selleck Chemicals (Cat. No. S7537, United States). A stock solution of LB-100 in 50 mM DNase/RNase-Free Distilled Water (Cat. No. 10977015, Thermo Fisher, United States) stored at −80°C was diluted as needed in cold phosphate buffered saline (PBS; Biosharp, China).

Rabbit monoclonal antibody (mAb) against YAP (Cat No. 52771), rabbit mAb against Ki67 (Cat No. 15580), mouse mAb against human CD31 (Cat No. 9498), rabbit mAb against ERG (Cat No. 92513) and rat mAb against BrdU (Cat No. 6326) were purchased from Abcam (Cambridge, United Kingdom). Rabbit mAb against YAP (Cat No. 14074), Rabbit mAb against pYAP (Cat No. 4911) and rabbit mAb against MST1 (Cat No. 14946) were from Cell Signaling Technology (Boston, MA, United States). Mouse mAb against β-actin, Mouse mAb against GAPDH and Mouse mAb against β-tubulin were from Utibody (Tianjin, China). LB100 (Cat No. s7537) was from Selleck (Houston, TX, United States). LatB (Cat No. L5288) was from Sigma Aldrich (St. Louis, MO, United States). Recombinant Human VEGF (Cat No. 293-VE) was from R&D systems (Minneapolis, MN, United States).

VPA (Sigma-Aldrich, P4543-100G) and HU (Sigma-Aldrich, H8627-25G) were purchased from Sigma-Aldrich. Chk1 inhibitor (LY2603618, Selleckchem, S2626) and PP2A inhibitor (LB-100, Selleckchem, S7537) were purchased from Selleck Chemicals.

Mice used in airway inflammation studies were C57Bl/6J purchased from Jackson Laboratories. For primary CD4⁺ T cell cultures, total CD4⁺ T cells were isolated from the lymph nodes and spleens of either C57Bl6/J mice or FoxP3-EGFP mice (Stock 016961 from Jackson Laboratories) by negative magnetic selection with the MACS CD4⁺ T cell isolation kit (Miltenyi). Cells were cultured in RPMI (Cell Gro) supplemented with 10% FBS, 1% PSQ, 1% non-essential amino acids, 1% Sodium Pyruvate, and 10mM HEPES (Gibco). For long-term assays, cell culture plates were coated with antibodies to CD3 and CD28 (eBiosciences—clones 17A2 and 37.51, respectively) to promote activation and proliferation. Short-term stimulation assays were performed using soluble CD3 and CD28, cross-linked with anti-hamster IgG (Jackson Immunolabs, 107-005-142). Cultures were treated with calyculin A (Calbiochem), okadaic acid (Sigma) or LB100 (Selleckchem).

Myoblasts were cultured in 6-well cell culture plates and infected with sh-2A and sh-NC at multiplicity of infection (MOI) 50. The PP2A inhibitor LB-100 (Selleck, Shanghai, China) was added 24 h post infection of adenoviruses at a final concentration of 1 µM. Myoblasts were induced to myogenic differentiation for 6 days before microscopy and RT-PCR analysis.