Fig. 2: The activation of TRPML1 channel induces the release of lysosomal Fe2+, which in turn activates PHDs, resulting in suppression of macrophage IL1B transcription. | Nature Communications

Fig. 2: The activation of TRPML1 channel induces the release of lysosomal Fe2+, which in turn activates PHDs, resulting in suppression of macrophage IL1B transcription.

From: Lysosomes finely control macrophage inflammatory function via regulating the release of lysosomal Fe2+ through TRPML1 channel

Fig. 2: The activation of TRPML1 channel induces the release of lysosomal Fe2+, which in turn activates PHDs, resulting in suppression of macrophage IL1B transcription.The alternative text for this image may have been generated using AI.

a Relative IL1B mRNA levels in BMDMs under control, ML-SA5 (0.1 μM), ML-SA5 + DXZ (200 μM), ML-SA5 + 2, 2’-Bipyridyl (150 μM), ML-SA5 + DFX (200 μM), ML-SA5 + Bapta-am (20 μM), ML-SA5 + TPEN (10 μM), or ML-SA5 + Wortmannin (100 nM pretreatment for 2 h). All treatments were for 24 h. n = 4-16. b Representative immunoblots and quantitative analysis of pro IL-1β protein levels in BMDMs under control, LPS, LPS + ML-SA5, LPS + ML-SA5 + DXZ, LPS + ML-SA5 + 2, 2’-Bipyridyl, LPS + ML-SA5 + DFX, DXZ, 2, 2’-Bipyridyl, or DFX. All treatments were for 24 h. n = 3–6. c Relative ODD-luciferase activity of BMDMs following control, ML-SA5 (0.1 μM), LPS (100 ng/mL), and LPS + ML-SA5 conditions was measured by ODD-luciferase reporter assays. All treatment were for 4 h. n = 4–8. d Relative ODD-luciferase activity of BMDMs following LPS, LPS + ML-SA5, LPS + ML-SA5 + ML-SI3 (10 μM), or LPS + ML-SA5 + 2, 2’-Bipyridyl conditions were measured by ODD-luciferase assays. All treatment were for 4 h. n = 4–8. e Relative IL1B mRNA levels in BMDMs under control, ML-SA5, ML-SA5 + Roxadustat (100 μM), ML-SA5 + Vadadustat (200 μM), or ML-SA5 + GSK1278863 (250 μM). All treatments were for 24 h. n = 4–10. f Representative immunoblots and quantitative analysis of pro IL-1β protein levels in BMDMs under control, LPS, LPS + ML-SA5, LPS + ML-SA5 + Roxadustat, LPS + ML-SA5 + Vadadustat, LPS + ML-SA5 + GSK1278863, Roxadustat, Vadadustat, or GSK1278863. All treatments were for 24 h. n = 3–6. g Relative IL1B mRNA levels under control, ML-SA5, LPS, or LPS + ML-SA5 in NC shRNA, PHD1 shRNA, PHD2 shRNA, PHD3 shRNA, or PHD1 shRNA + PHD2 shRNA (denoted as PHD1 + 2 shRNA) expressed BMDMs. All treatments were for 24 h. n = 4–6. h Representative immunoblots and quantitative analysis of pro IL-1β protein levels under LPS or LPS + ML-SA5 in NC shRNA, PHD1 shRNA, PHD2 shRNA, PHD3 shRNA, or PHD1 + 2 shRNA expressed BMDMs. All treatments were for 24 h. n = 3–12. Means ± SEMs were shown in a–h. Significant differences were evaluated using one-way ANOVA followed by Tukey’s test.

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