![Figure 4. NF-κB activity in response to TNF-α and IL-1β. (A) IκBα degradation, detected by Western blotting and induced by TNF-α and IL-1β, for a healthy control subject (C+), an XD-IP patient, a hypomorphic NEMO EDA-ID-OL patient (X420W), the proband (P [D311G]), and an XR-MSMD NEMO patient (R319Q). Antibodies against GAPDH were used for normalization of protein levels on the Western blot. (B) NF-κB translocation was observed by EMSA with a labeled probe after exposure to TNF-α and IL-1β for various periods of time (0, 20, 40 minutes) for a healthy control subject (C+), an XD-IP patient, a hypomorphic NEMO EDA-ID-OL patient (X420W), the proband (P [D311G]), and an NEMO XR-MSMD patient (R319Q). (C) IL-10 production after TNF-α activation in whole blood from healthy control subjects, the proband (P [D311G]), and an NEMO XR-MSMD patient (R319Q). (D) IL-6 and IL-8 production, determined by ELISA after 24 hours of activation with TNF-α and IL-1β, for SV40-transformed fibroblast cell lines from a healthy control subject (C+), an XD-IP patient, an NEMO EDA-ID-OL patient (X420W), the proband (P [D311G]), and an NEMO XR-MSMD patient (R319Q). These results are representative of 2 independent experiments.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/118/4/10.1182_blood-2010-10-315234/4/zh89991174700004.jpeg?Expires=1736058914&Signature=F928V--gzGeXWFPbZjTISG6pUzYdooKj7mYipHlBDe88pDR7-~u6h08NeFLLzGdRRXDxxEMb8P3W5fT8lUUGsoXmZHAoTcquPYAxJ69KDB6zhs0aLYJQUef1lPExME~5PaQojsaRVba08Eq9XsB~WsvVewSfgtS3Te3~JHheiI542AmPhyNPh~8X5Rc0Dc0qxMF3wIEvzCjCF~tOd92dOkN~L-WaIa0VwKgrHpuM1BxYbN2GPzvklkJcNjJTFFkMQ-6Z111jt55Iir-dDJzMhqFEKSTgtEmeWONfLa-MOuJIzDJqNKJVRgPdfW7nz8vIB630UmzfwrlyuSPU-h4h0w__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
NF-κB activity in response to TNF-α and IL-1β. (A) IκBα degradation, detected by Western blotting and induced by TNF-α and IL-1β, for a healthy control subject (C+), an XD-IP patient, a hypomorphic NEMO EDA-ID-OL patient (X420W), the proband (P [D311G]), and an XR-MSMD NEMO patient (R319Q). Antibodies against GAPDH were used for normalization of protein levels on the Western blot. (B) NF-κB translocation was observed by EMSA with a labeled probe after exposure to TNF-α and IL-1β for various periods of time (0, 20, 40 minutes) for a healthy control subject (C+), an XD-IP patient, a hypomorphic NEMO EDA-ID-OL patient (X420W), the proband (P [D311G]), and an NEMO XR-MSMD patient (R319Q). (C) IL-10 production after TNF-α activation in whole blood from healthy control subjects, the proband (P [D311G]), and an NEMO XR-MSMD patient (R319Q). (D) IL-6 and IL-8 production, determined by ELISA after 24 hours of activation with TNF-α and IL-1β, for SV40-transformed fibroblast cell lines from a healthy control subject (C+), an XD-IP patient, an NEMO EDA-ID-OL patient (X420W), the proband (P [D311G]), and an NEMO XR-MSMD patient (R319Q). These results are representative of 2 independent experiments.
