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. 2019 Apr 30;294(24):9503–9517. doi: 10.1074/jbc.RA118.006563

Figure 5.

Figure 5.

Mapping of the STAT2 domains required for efficient HNSs–STAT2 interaction. A, domain organization of full-length or N-terminal truncated STAT2 C-terminally fused with HA tag. NTD, N-terminal domain; CCD, coiled-coil domain; SH2, src homology domain-2; TAD, transactivation domain; pY, tyrosine (Tyr-690) phosphorylation site. The HA-tagged truncated STAT2 proteins were named T1-HA, T2-HA, T3-HA, T4-HA, and T5-HA, respectively. B, HEK293 cells were transfected with the HNSs-S expression plasmid and the plasmids encoding full-length or truncated STAT2 proteins or the corresponding control vectors, as indicated. At 48 h posttransfection, interactions of HNSs with the full-length or truncated STAT2 were analyzed with the S-pulldown assay, followed by WB with the indicated antibodies. C, band intensities of full-length or truncated STAT2 proteins in B were respectively measured by ImageJ software. To calculate the pulldown ratio of full-length or truncated STAT2, band intensities of the proteins co-precipitated with HNSs were then normalized to the corresponding band intensities in lysate input. The relative pulldown ratio of full-length STAT2 was set to 1, for reference. N.A., not analyzed.