Pma tpa

BCBL-1 cells were grown in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS, Gibco-BRL, Gaithersburg, MD, USA). For the lytic replication of KSHV, BCBL-1 cells were induced with 20 ng/mL 12-O-tetradecanoyl-phorbol-13-acetate (PMA/TPA) (Beyotime Institute of Biotechnology, Wuhan, China). Human iSLK.219 cells, derived from iSLK cells, were latently infected with a recombinant rKSHV.219 virus and also contained a doxycycline-inducible RTA [26 (link)]. The rKSHV.219 expresses the red fluorescent protein (RFP) from the KSHV lytic PAN promoter, the green fluorescent protein (GFP) from the EF-1α promoter [27 (link)]. Human iSLK.219 cells were cultured in DMEM supplemented with 10% FBS. The cells were reactivated with 1 µg/mL doxycycline (Dox) and 1.2 mM sodium butyrate (NaB). The chemicals used for cell culture were purchased from Invivogen (San Diego, CA, USA). Celecoxib was purchased from Sigma-Aldrich (St. Louis, MO, USA), NS-398, SP600125 and SB203580 were purchased from Beyotime Institute of Biotechnology, and nimesulide and cidofovir (CDV) were obtained from Selleck Chemicals (Shanghai, China). All drugs were dissolved in dimethylsulfoxide (DMSO).

Rabbit monoclonal CXCR4 antibody (ab124824), rabbit monoclonal LC3B antibody (ab192890), rabbit monoclonal NRF1 antibody (ab175932), and rabbit monoclonal ZEB1 antibody (ab203829) were purchased from Abcam (UK). Rabbit anti-beta-actin (β-actin) antibody (#4970), rabbit anti-AKT antibody (#4685), and rabbit anti-p-AKT antibody (Ser473, #4060) were purchased from CST (USA). Anti-EBV BZLF1 antibody (sc-53904) was purchased from Santa Cruz Biotechnology (USA). Anti-rabbit and anti-mouse secondary antibodies were purchased from Abcam. Goat anti-rabbit polyclonal secondary antibody conjugated with Alexa Fluor 647 (ab150079) was purchased from Abcam. Anti-rabbit antibody conjugated with Alexa Fluor 555 (#4413) and anti-mouse antibodies conjugated with Alexa Fluor 555 (#4409) were purchased from CST. All antibodies were used according to the manufacturer's instructions. The PI3K/AKT signalling pathway inhibitor LY294002 and EBV reactivation inducer PMA/TPA were purchased from Beyotime Biotechnology (China). Rapamycin (autophagy inducer) was purchased from Apexbio (USA).