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. 2023 May 1;53(7):2250056. doi: 10.1002/eji.202250056

Figure 7.

Figure 7

Role of SCIMP and SYK in TLR4 Y672‐dependent cytokine responses. (A‐F) Tlr4 −/− BMM were reconstituted with WT mouse TLR4, TLR4 mutants or empty vector (EV). (A‐C) Cells were electroporated with Scimp siRNA, a control siRNA or no siRNA, after which cells were treated with LPS (10 ng/mL) for 24 h. Supernatants were collected and assessed for levels of IL‐6, IL‐12p40, and TNF by ELISA. Dashed lines indicate cytokine levels in Y672F‐expressing BMM transfected with control siRNA to facilitate comparison between conditions. Data (mean + SEM, n = 5) are combined from five independent experiments with different symbols showing data points from each experiment and are normalized to the control siRNA WT TLR4 sample. (D‐F) Transduced cells were preincubated with 10 μM of the SYK‐specific inhibitor SykIV for 30 min, then stimulated with LPS (10 ng/mL) for 24 h. Supernatants were collected and assessed for levels of IL‐6, IL‐12p40, and TNF by ELISA. Data (mean + SEM, n = 3) are combined from three independent experiments and are normalized to the WT TLR4 DMSO + LPS‐treated sample. (A‐F) Statistical analyses were performed via a repeated measures two‐way ANOVA, followed by Bonferroni's multiple comparison test (NS: non‐significant, *p < 0.05, **p < 0.01, ****p < 0.0001).