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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Cancer-derived exosomal TRIM59 regulates macrophage NLRP3 inflammasome activation to promote lung cancer progression

Fig. 4

ABHD5 deficiency promotes NLRP3 inflammasome activation in macrophages. a. Western blot analysis of ABHD5 expression in macrophages transfected with scrambled control shRNA or ABHD5 shRNA for 36 h. Immunoblot of the IL-1β, the pro-caspase-1 and cleaved caspase-1 in the supernatants (SNs) or cell lysates of ABHD5-silenced THP-1 macrophages, primed with LPS, and then stimulated with Nigericin (Nig.), ATP or Alum. β-actin served as a loading control. Quantification of Western blotting were performed with the Image J software. Numbers below each blot indicate relative band intensity normalized to β-actin. b. ELISA of IL-1β in supernatants from THP-1 macrophages silenced of ABHD5, primed with LPS for 8 h, and followed by stimulation with ATP, Nig., Alum, poly(dA:dT) or flagellin for 30 min. c. ELISA of IL-1β in supernatants from BMDMs silenced of ABHD5, primed with LPS for 8 h, and followed by stimulation with ATP, Nig., Alum, poly(dA:dT) or flagellin for 30 min. d. RT-PCR analysis of IL-1β mRNA expression in macrophages transfected with shRNA as indicated and stimulated as indicated. e. ELISA of IL-1β in supernatants from THP-1 cells infected with lentiviral vectors expressing ABHD5 or GFP control, primed with LPS for various times, and followed by stimulation with ATP for 30 min. f. ELISA of IL-1β in supernatants from THP-1 cells infected with lentiviral vectors expressing TRIM59 or GFP control, primed with LPS, and followed by stimulation with ATP or Nig. Data are from three independent experiments with biological duplicates in each (mean ± SEM) or are representative of three independent experiments. Student’s t-test, **p < 0.01

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