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. Author manuscript; available in PMC: 2023 Nov 8.
Published in final edited form as: Immunity. 2022 Oct 14;55(11):2074–2084.e5. doi: 10.1016/j.immuni.2022.09.007

Figure 2. Initial hyper-response to IFN-I underlies heightened IFN-I negative regulation.

Figure 2.

(A) Immunoblotting for STAT phosphorylation after stimulation for 15 min with IFN-α (100 IU/mL) of HC and DS hTERT-immortalized fibroblasts previously stimulated (Primed) or not (Naive) with a primary stimulus of IFN-α (10 IU/mL) for 12 h, washed, and allowed to rest for 36 h. Result representative of fibroblasts derived from n=3 HCs and n=3 individuals with DS.

(B) Immunoblotting for STAT phosphorylation after stimulation for 15 min with IFN-α (100 IU/mL) of IFNAR2 knockouts complemented with doxycycline-inducible IFNAR2, treated with indicated amounts of doxycycline and previously stimulated (Primed) or not (Naïve) with a primary stimulus of IFN-α (10 or 100 IU/mL). Results representative of n=3 independent experiments.

(C) Immunoblotting for STAT phosphorylation after stimulation for 15 min with IFN-α (100 IU/mL) of IFNAR2 knockouts complemented with doxycycline-inducible IFNAR2, treated with indicated amounts of doxycycline and previously stimulated (Primed) or not (Naïve) with a primary stimulus of IFN-α (10 or 100 IU/mL).

(D) Immunoblotting for STAT phosphorylation after stimulation for 15 min with IFN-α (100 IU/mL) of IFNAR2 knockouts complemented with doxycycline-inducible IFNAR2, treated with increasing concentrations of doxycycline and Primed (with 10 IU/mL IFN-α) or not (Naïve), in the presence (parental line) or absence (USP18 KO Clones 1 and 2) of endogenous USP18.

(E) Diagram of hyper- and hypo-responses to IFN-I over time with subsequent stimulations in HC and DS individuals.

See also Figure S3.