Fig. 5: TPL2 mediates JNK activation by LMP1, but it is dispensable for NF-κB induction.

a The knockdown of TPL2 interferes with JNK activation by LMP1. Wildtype MEF:NGFR-LMP1wt cells were transfected twice with siRNA targeting TPL2 (siTPL2) or non-targeting control siRNA (siCTRL) prior to NGFR-LMP1 activation. b Pharmacological inhibition of TPL2 attenuates JNK activation by LMP1. Detection of JNK and NF-κB activation by NGFR-LMP1 in wtMEF:NGFR-LMP1wt cells in the presence of DMSO or 10 μM of the TPL2 inhibitor TC-S7006 (TPL2-IH). c The knockout of TPL2 inhibits JNK signaling by LMP1. TPL2 was targeted by CRISPR/Cas9 in wtMEF:NGFR-LMP1wt cells. NGFR-LMP1 was induced in wildtype and TPL2^crKO (clone 8) MEFs. JNK and NF-κB activation were analysed. d The knockout of TPL2 in HEK293 cells does not impair canonical or non-canonical NF-κB activation by LMP1. TPL2 was inactivated in HEK293 cells by CRISPR/Cas9 technology. HEK293 wildtype and TPL2^crKOHEK293 (clone 1) cells were transfected with HA-LMP1 wildtype, inactive AAA/Δ371–386 mutant or empty vector as indicated. Cytoplasmic and nuclear fractions were prepared and analysed by immunoblotting with the indicated antibodies. TPL2 knockouts were verified at the protein level (this Figure) and by sequencing (Supplementary Table 2). a–d The data are representative of at least two independent experiments. For immunoblot quantification and statistics see Supplementary Table 3.