Figure 2.
XIAP-deficient cells produce excess IL-1β when stimulated under serum starvation. (A) Monocytes were isolated from PBMC obtained from the proband and his healthy father and stimulated with media alone (unstimulated), LPS, or LPS with ATP added for the last hour of incubation. After 4 hours, supernatants were taken for IL-1β measurement using enzyme-linked immunosorbent assay (ELISA). (B) Control or XIAP-KO THP-1 macrophages were stimulated as indicated in panel A, followed by supernatant retrieval for IL-1β measurement using ELISA. (C) Control or XIAP-KO THP-1 macrophages were stimulated with media alone (unstimulated), LPS, or LPS with ATP added for the last hour of incubation. After 24 hours, supernatants were taken for IL-1β measurement using ELISA. (D) Monocytes isolated from PBMC obtained from the proband and his healthy father were stimulated as indicated in panel A under standard (10% FCS) or starvation (1% FCS) conditions, after which supernatants were taken for IL-1β measurement by ELISA. Panels A-D are representative of >3 independent experiments and indicate mean with standard deviation. (E) Control or XIAP-KO THP-1 macrophages were stimulated with LPS or LPS with ATP added for the last hour of incubation in serum starvation (1% FCS) media for 4 hours, after which supernatants were harvested for IL-1β measurement using ELISA. (F) Supernatants obtained as in panel E were subjected to ELISA for TNF-α measurement. For panels E-F, results are pooled from 3 independent experiments and normalized using measurements from supernatants of cells incubated with the same stimuli under nonserum starvation conditions. The median value is indicated with a line, the box indicates 25th to 75th percentile, and the whiskers indicate the minimum and maximum values.

XIAP-deficient cells produce excess IL-1β when stimulated under serum starvation. (A) Monocytes were isolated from PBMC obtained from the proband and his healthy father and stimulated with media alone (unstimulated), LPS, or LPS with ATP added for the last hour of incubation. After 4 hours, supernatants were taken for IL-1β measurement using enzyme-linked immunosorbent assay (ELISA). (B) Control or XIAP-KO THP-1 macrophages were stimulated as indicated in panel A, followed by supernatant retrieval for IL-1β measurement using ELISA. (C) Control or XIAP-KO THP-1 macrophages were stimulated with media alone (unstimulated), LPS, or LPS with ATP added for the last hour of incubation. After 24 hours, supernatants were taken for IL-1β measurement using ELISA. (D) Monocytes isolated from PBMC obtained from the proband and his healthy father were stimulated as indicated in panel A under standard (10% FCS) or starvation (1% FCS) conditions, after which supernatants were taken for IL-1β measurement by ELISA. Panels A-D are representative of >3 independent experiments and indicate mean with standard deviation. (E) Control or XIAP-KO THP-1 macrophages were stimulated with LPS or LPS with ATP added for the last hour of incubation in serum starvation (1% FCS) media for 4 hours, after which supernatants were harvested for IL-1β measurement using ELISA. (F) Supernatants obtained as in panel E were subjected to ELISA for TNF-α measurement. For panels E-F, results are pooled from 3 independent experiments and normalized using measurements from supernatants of cells incubated with the same stimuli under nonserum starvation conditions. The median value is indicated with a line, the box indicates 25th to 75th percentile, and the whiskers indicate the minimum and maximum values.

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