Stempro msc sfm basal medium cts

H290 cells cultured in the flask to 80% confluence were trypsinized to prepare single‐cell suspensions (one cell per μL), which were treated with 100 µmol/L THZ1 (a Cdk7 inhibitor) (CAS# 1604810‐83‐4, Cayman CHEMICAL) or DMSO (0.05%) for 24 hours in StemPro MSC SFM Basal Medium CTS + StemPro MSC SFM Supplement CTS (Life Technologies), 2 nmol/L l‐glutamine and penicillin (100 IU/mL). Then, 2 × 103 cells per well were plated in an ultra‐low 12‐well attachment plates (Corning Incorporated). Cells were cultured for 10 days. Then, 100 μL of medium was added into each well every 2‐3 days. Tumorspheres that formed in non‐adherent cultures were counted under a 10× objective lens. The cut‐off size for the spheres counted was 60 µm.

One cell per μl H290 single‐cell suspensions after treatment with control siRNA or YAP siRNA‐1 for 48 hrs or after treatment with 0.3 μM verteporfin, 1.0 μM verteporfin or DMSO (0.1%) for 24 hrs were prepared in StemPro MSC SFM Basal Medium CTS + StemPro MSC SFM Supplement CTS (Life Technologies), 2 nM L‐glutamine and penicillin (100 IU/ml). Then, 200 μl (200 cells per well) of the cells were plated in 96‐well ultra‐low attachment plates (Corning Incorporated). For each treatment, seed cells into the wells of two rows for a total of 20 wells. Tumoursphere were cultured for 7 days. Tumoursphere formed in non‐adherent cultures were counted under a 10× objective lens. The cut‐off size for the spheres counted was 60 μm.