Journal: Chinese Medicine

Article Title: Ivangustin, a sesquiterpene lactone from Inula japonica Thunb., attenuates neuroinflammation through covalent inhibition of ubiquitin-conjugating enzyme UBE2L3

doi: 10.1186/s13020-026-01488-9

Figure Lengend Snippet: The effects of IVA on LPS-induced activation of NF-κB pathway in BV2 cells. Western blotting analysis of p-IKKα/β and total IKKα/β ( A ), p-IκBα and total IκBα ( C ), as well as p-p65 and total p65 ( E ) in BV2 cells following IVA treatment (2 h) and LPS stimulation (5 min). Densitometric quantification of p-IKKα/β, p-IκBα, IκBα, and p-p65, normalized to their total forms or Tubulin, respectively, is presented in ( B , D , and F ). For NF-κB p65 subcellular localization analysis, cytoplasmic and nuclear fractions from BV2 cells treated with IVA (2 h) and stimulated with LPS (30 min) were subjected to immunoblotting ( G ), with GAPDH and PARP1 serving as loading controls for cytoplasmic and nuclear fractions, respectively. Statistical results of p65 distribution, normalized to GAPDH or PARP1, are shown in ( H ). BV2 cells were treated with IVA (5, 10, and 20 μM), SP600125 (SP, 20 μM), or SB203580 (SB, 20 μM) for 2 h, followed by LPS stimulation for 30 min. I The phosphorylation levels of JNK and p38, as well as total protein levels of JNK, p38, GAPDH, and Tubulin were detected by western blotting (SP and SB were used as positive controls for p-JNK and p-p38, respectively). J The quantitative results of p-JNK and p-p38 normalized against total JNK and p38 are shown, respectively. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, vs. LPS; ns means no significant; n = 3

Article Snippet: Dimethyl fumarate (DMF), SP600125 (SP, 20 μM, a selective JNK inhibitor), and SB203580 (SB, 20 μM, a selective p38 inhibitor) were obtained from TargetMol (Shanghai, China).

Techniques: Activation Assay, Western Blot, Analysis, Phospho-proteomics