OTULIN deficiency in ORAS causes cell type‐specific LUBAC degradation, dysregulated TNF signalling and cell death

A

Immunoblot analysis of whole‐cell lysate from untreated human THP‐1 monocytes with stable expression of a non‐targeting control shRNA or and shRNA targeting OTULIN. Data are representative of two independent experiments.

B

Immunoblot (left) and densitometry (right) analysis of IκBα levels in shControl and shOTULIN THP‐1 cells treated with cycloheximide (CHX) (50 μg/ml) as indicated. Data are representative of three independent experiments.

C

ELISA analysis of spontaneous TNF secretion over 96 h in shControl and shOTULIN THP‐1 cells. Data represent mean ± SEM (n = 4) and were analysed using the two‐way ANOVA test of statistical significance with Sidak's correction for multiple comparisons.

D

Immunoblot analysis of IκBα phosphorylation and degradation as well as p65/RelA phosphorylation in shControl and shOTULIN THP‐1 cells in response to stimulation with TNF (10 ng/ml). Data are representative of three independent experiments.

E

Densitometry analysis of p65/RelA phosphorylation as presented in (D) from three independent experiments.

Figure 5. OTULIN deficiency in human THP‐1 monocytes leads to increased NF‐κB activation and spontaneous TNF secretion.