Defined fbs

Human CML cell line K562 was obtained from the American Type Culture Collection (ATCC, Rockville, MD). K562/ADR, an adriamycin–resistant cell line, and K562/G, an imatinib–resistant cell line, were kindly provided by Dr. Wei He (Institute of Hematology, The First Affiliated Hospital, College of Medicine, Zhejiang University, China). All cells were maintained in RPMI‐1640 medium (Hyclone Laboratories) supplemented with 10% defined FBS (Hyclone Laboratories). Cells were cultured at 37°C in 5% CO₂. Panobinostat (LBH589) was purchased from Selleck Chemicals (Houston, TX). Sodium butyrate (NaBu) was purchased from Sigma‐Aldrich (St. Louis, MO). The primary antibodies against β‐ACTIN, GAPDH, caspase‐3, caspase‐6, caspase‐8, caspase‐9, PARP, BIP, BCL‐XL, MCL‐1, BAX, BAK, BAD, XIAP, survivin, ABCG2, MDR1, BCR/ABL, p‐BCR/ABL, AKT, p‐AKT, mTOR, p‐mTORC1, 4EBP1, p‐4EBP1, eIF4E, p‐eIF4E and c‐MYC were purchased from Cell Signaling Technology (Danvers, MA), the antibody against caspase‐7 was purchased from Abcam (Cambridge, UK).

BMDMs were prepared as described previously [76 (link)]. On day 7, BMDMs were plated in 96 well plate (Corning) at a cell density of 0.08 x 10⁶ cells/well in 200μL of D10 media [antibiotic free DMEM media (Mediatech, Inc.) containing 10% defined FBS (HyClone Laboratories, Logan, UT)] and supplemented with 2% conditioned medium from L-cells. Cells were infected in replicates of 5 with 3 MOI of three Mtb-HT and three Mtb-LT strains for 4 hours. Wells were then washed 4 times with PBS + 1% BCS and cells were untreated or treated with 100nM of MPN. Infected cells were maintained in D-10 media supplemented with 2% conditioned medium from L-cells. At day 7, post-infection cells were washed with serum-containing PBS and then lysed with sterile water. Total CFU was determined by plating 10-fold serial dilutions on Middlebrook 7H11 plates, which were counted after 21 days of incubation at 37 °C. 48-hour culture supernatants were harvested for measuring TNF levels.

The human HCC cell line MHCC97-H was provided by Dr. Xinwei Wang, from the National Cancer Institute (NCI), under agreement with the Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, China, and was cultured as previously described [11 (link),29 (link)]. The human HCC cell lines Huh7 and Hep3B [acquired from AddexBio (San Diego, CA)] were maintained as previously described [30 (link)]. The human SK-Hep1 cells were provided by Dr. Brian Barth, Penn State College of Medicine, and maintained in Dulbecco’s modified Eagles Medium 1X supplemented with 10% defined FBS (Hyclone Laboratories, Logan, UT), 1 mM GlutaMAX-1 (Life Technologies), 100 U/ml penicillin and 100 μg/ml streptomycin. The cells were cultured in a humidified incubator with 5% CO₂ at 37°C.