Fig. 7: PB2s of H5 subtype AIVs degrade mammalian JAK1 differently.
From: The influenza virus PB2 protein evades antiviral innate immunity by inhibiting JAK1/STAT signalling
a Immunoblots of HEK293T cells transfected with JAK1 and PB2-CZ, PB2M-CZ (M283I-R526K), PB2-JY, or PB2M-JY (I283M-K526R) plasmids (upper). The intensities of the bands on the immunoblots from three independent experiments were quantified and normalized with actin (lower). Data are presented as the mean ± SD. b Immunoblots of HEK293T cells transfected with JAK1 and PB2-CZ, PB2-R526K-CZ, PB2-M283I-CZ, or PB2M-CZ plasmids (upper). The intensities of the bands on the immunoblots from three independent experiments were quantified and normalized with actin (lower). Data are presented as the mean ± SD. c Co-ip analysis of the interaction of PB2 or its mutants with JAK1 in HEK293T cells. d Colocalization of endogenous JAK1 (green) and PB2 (red) in rJY, rCZM (M283I-R526K), or rJYM (I283M-K526R) infected A549 cells. Nuclei were stained with DAPI (blue). Scale bars, 10 μm. Intensities of fluorescence at indicated locations were scanned by LAS X Software. e Ni-NTA pull-down analysis of the ubiquitination of JAK1 in HEK293T cells transfected with PB2 or its mutant plasmids and treated with MG132. WCL, whole-cell lysates. Statistical significance in a, b was determined by unpaired two-tailed Student’s t test. ns P > 0.05. Data are one representative of three independent experiments.
