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. 2015 Dec 30;90(2):1129–1138. doi: 10.1128/JVI.01410-15

FIG 2.

FIG 2

Expression of BGLF2 induces p38 and JNK phosphorylation. (A) 293T cells were transfected with a plasmid expressing the BGLF2 protein with a V5 epitope tag, a plasmid expressing MEKK1 (positive control), or pcDNA3.1 (negative control), and after 48 h, cell lysates were immunoblotted with an antibody to p38, p-p38, JNK, p-JNK, Akt, p-Akt, ERK1/2, p-ERK1/2, actin, or V5 (to detect BGLF2). (B) 293T-BGLF2 cells, which express BGLF2 under the control of a doxycycline (DOX)-inducible promoter, were treated with or without DOX for 2 days to induce expression of BGLF2, and cell lysates were immunoblotted for signaling proteins. (C) 293T-lenti6-BGLF2 cells, which constitutively express BGLF2, were transduced with a lentivirus-mediated shRNA against BGLF2 (shBGLF2) or luciferase (shLuc), and after 4 days, cell lysates were collected for immunoblotting. (D) Akata-EBV cells were transduced with a lentivirus expressing BGLF2 under the control of a DOX-inducible promoter and were treated with or without DOX for 4 days, and cell lysates were collected to detect p-p38, p38, p-JNK, JNK, BGLF2 (using an antibody to V5), and actin.