Fig. 10: NRF2 is essential for TSC1-mediated immune regulation in macrophages in vitro. | Cell Death & Disease

Fig. 10: NRF2 is essential for TSC1-mediated immune regulation in macrophages in vitro.

From: Loss of macrophage TSC1 exacerbates sterile inflammatory liver injury through inhibiting the AKT/MST1/NRF2 signaling pathway

Fig. 10

Bone marrow-derived macrophages (BMMs) were isolated from NRF2FL/FL and NRF2M-KO mice and pretreated with Lv-TSC1 or Lv-GFP controls before LPS stimulation for 6 h. A ROS production was detected by Carboxy-H2DFFDA in LPS-stimulated BMMs from TSC1M-KO mice, Scale bars = 20 μm. B Quantification of ROS-producing BMMs (green). **p < 0.01. C Western blot analysis of TSC1, NRF2, HMGB1, NF-κB and TLR4. β-actin served as an internal control. Data are representative of three experiments. D Quantitative RT-PCR-assisted detection of TNF-α, IL-1β, IL-6 and TGF-β in LPS-stimulated BMMs. Mean ± SD (n = 3–4 samples/group). *p < 0.05, **p < 0.01. E BMMs were transfected with CRISPR-TSC1 activation vector (1 and 3 µg, respectively). Immunoprecipitation analysis of Keap1 and NRF2 in macrophages after LPS stimulation. F Immunoblot analysis of NRF2 ubiquitination in total lysates (bottom) and anti-IgG or anti-NRF2 immunoprecipitates (IP, top) of WT and TSC1-KO BMMs, probed with anti-ubiquitin (α-Ub) and antibodies to NRF2, TSC1, and GAPDH.

Back to article page