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. 2021 Jan 13;95(3):e01970-20. doi: 10.1128/JVI.01970-20

FIG 4.

FIG 4

SAMHD1 inhibits NF-κB activation by TAB3/TAK1 or TRAF2. (A) HEK293T cells were cotransfected with the indicated amounts of pRK-HA-TAB3, pRK-HA-TAK1, wild-type pCG-F-HA-SAMHD1 (pSAMHD1), 50 ng pNF-κB-luciferase, and 10 ng of TK-renilla. Immunoblotting was performed with specific antibodies to the indicated proteins. (B) HEK293T cells were cotransfected with the indicated amounts of, pcDNA3.1-F-TRAF2, pCG-F-HA-SAMHD1 (pSAMHD1), 50 ng pNF-κB-luciferase, and 10 ng of TK-renilla. The total amount of DNA was maintained through the addition of empty vector (V). Cell lysates were collected for the luciferase assay and immunoblotting at 24 h posttransfection. All luciferase values were normalized to 10 μg protein. Relative NF-κB activity was calculated by setting empty vector-transfected cells as 1. Immunoblotting of SAMHD1 and TRAF2 was performed with anti-FLAG antibodies. GAPDH was used as a loading control. Statistical analysis was performed by unpaired t test with Welch’s correction. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.