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. 2013 Apr 11;50(1):149–156. doi: 10.1016/j.molcel.2013.02.024

Figure 2.

Figure 2

β-Arrestin1 Does Not Affect STAT1 Phosphorylation

(A) Immunoblot showing a time course of STAT1 phosphorylation in IFNγ-treated HeLa cells that transiently express β-arrestin1 or β-galactosidase as indicated. Cells were left untreated or treated with IFNγ for 60 min, followed by incubation for 0–240 min in growth medium (GM) without interferon. Probing was done first with anti-Tyr701-phoshorylated STAT1 antibody, then with a mixture of anti-human STAT1 and anti-β-actin antibody, followed by a mixture of anti-GFP and anti-FLAG-tag, and finally with anti-β-galactosidase. Molecular weight marker positions are indicated. The diagram combines the results of this and another experiment. It shows specific Tyr701-phosphorylation of STAT1 under the different experimental conditions (specific Tyr701 phosphorylation after 60 min IFNγ was set as 100). Results were calculated as the mean ± standard deviation.

(B) Shown are immunoblot analyses of wild-type (WT) and β-arrestin1-deficient MEFs using anti-β-arrestin1 antibody. Reprobing was done with anti-β-actin. Positions of molecular weight markers and β-arrestin 1 are indicated.

(C) Top, immunoblot analyses of STAT1 dephosphorylation in WT and mutant MEFs. Bottom, diagram combining the results of this and two additional experiments giving specific Tyr701 phosphorylation of STAT1. The value for 60 min IFNγ was set as 100. Results were calculated as the mean ± standard deviation.