nf κb inhibitor mcck1  (MedChemExpress)

Journal: International Journal of Molecular Sciences

Article Title: Aberrant CX3CL1-CX3CR1 Signaling Reprograms Microglial Exosome Secretion via KIFC2 to Drive Cognitive Impairment in Chronic Pain

doi: 10.3390/ijms27146304

Figure Lengend Snippet: The microglial CX3CL1-CX3CR1 signaling induces neurotoxicity by promoting IL-17-enriched exosome release via the p38 MAPK-NF-κB-KIFC2 axis. ( A , B ) Western blot analysis of p38 MAPK-NF-κB phosphorylation and KIFC2 expression in BV2 cells treated with CX3CL1, in the presence of the p38 inhibitor SB203580 (20 μM) or the NF-κB inhibitor MCCK1 (2 μM). ( C – F ) NTA characterization of EVs secreted by BV2 cells treated with CX3CL1 in combination with SB203580 or MCCK1, displaying representative size distribution profiles ( C ), absolute concentration of small exosomes (<100 nm) ( D ), total EV concentration ( E ), and the relative proportion of exosomes ( F ). ( G ) ELISA quantification of IL-17 concentrations in EVs secreted by BV2 cells following the indicated inhibitor treatments. ( H ) CCK-8 assay assessing the viability of PC12 cells co-cultured with BV2 cells pre-treated with CX3CL1, SB203580, or MCCK1. ( I ) Western blot analysis of PSD95 and cleaved caspase-3 (c-Casp3) expression in PC12 cells co-cultured with similarly pre-treated BV2 cells. p -values were determined by one-way ANOVA followed by Tukey’s post hoc test ( D – H ).

Article Snippet: For pharmacological inhibition, BV2 cells were pre-incubated with either the p38 inhibitor SB203580 (HY-10256, MedChemExpress, 20 μM) or the NF-κB inhibitor MCCK1 (HY-D0162R, MedChemExpress, 2 μM) for 12 h, followed by stimulation with recombinant murine CX3CL1 (RP01393, ABclonal, Woburn, MA, USA) for 24 h. For gene silencing experiments, BV2 cells were transfected with small interfering RNAs (siRNAs) targeting Kifc2 or Cx3cr1 for 36 h, followed by a 24 h stimulation with recombinant CX3CL1.

Techniques: Western Blot, Phospho-proteomics, Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Cell Culture

Journal: Cell Insight

Article Title: Targeting the ICAM-1/LFA-1 adhesion axis attenuates lethal Crimean-Congo hemorrhagic fever virus infection

doi: 10.1016/j.cellin.2026.100326

Figure Lengend Snippet: NF-κB signaling pathway is involved in CCHFV-induced upregulation of ICAM-1 . (A) Detection of p65 nuclear translocation. HUVECs were infected with CCHFV-GFP and treated with TPCA-1 (0, 5, 10 μM) throughout infection. Immunostaining was performed with anti-p65 antibody (red). Green, CCHFV infected cells; blue, nuclei (Hoechst 33258). (B) Western blot was used to detect the expression of ICAM-1, CCHFV NP and phosphorylated p65. Mock, uninfected cells. β-actin, loading control. (C) qRT-PCR detection of ICAM1 transcript levels in cells with different treatments. The expression levels were standardized by GAPDH and compared with the mock control. Data are presented as the mean ± SD from three replicates. (D) Representative fluorescence microscopic images of adhesion experiments treated with TPCA-1. Green, CCHFV infection; Red, adherent Jurkat cells. (E) Adhesion level was calculated by normalizing to mock control group and the result represents the fold change of the average fluorescence area of the adherent Jurkat cells. The data were calculated using FIJI on ten fluorescence images from two replicates and expressed as means ± SD. All statistical analyses were performed using one-way ANOVA, followed by Tukey's multiple comparisons test. ∗∗∗∗ P < 0.000 1. Scale bar, 50 μm.

Article Snippet: The following experiment-specific modifications were applied to assess the impact of the various interventions: use of ICAM1 knockdown and corresponding control cells for adhesion assays; preincubation of Jurkat-RFP cells with lifitegrast (MCE, Cat. No. HY-19344) (0, 10, 50, 250 and 1 250 nM, 30 min) before co-culture; preincubation with sICAM-1 (0, 200, 400, 800 μg/mL, 30 min; BSA, control); and addition of NF-κB inhibitor TPCA-1 (MCE, Cat. No. HY-10074) (0, 5, 10 μM) during HUVECs infection.

Techniques: Translocation Assay, Infection, Immunostaining, Western Blot, Expressing, Control, Quantitative RT-PCR, Fluorescence

Journal: Nature Metabolism

Article Title: Tissue-specific fibroblast lipid cues impose the rate of epithelial cancer invasion

doi: 10.1038/s42255-026-01514-y

Figure Lengend Snippet: a , SM metabolic pathway and inhibitors targeting enzymes. b – d , Spheroid invasion of oSCC with control media (grey), 5 µM SM (blue) or 5 µM SM and 20 µM imipramine (yellow) ( b ), oral FB secretomes (grey), with 20 µM imipramine (blue) or 20 µM imipramine and 2 µM S1P (yellow) ( c ) and oral FB secretomes (grey), with 2 µM PF543 (blue) or 2 µM PF543 and 2 µM S1P ( d ) (two-sided Mann–Whitney U- test, n = 6 replicates, two cell lines, two independent experiments). e , S1P immunofluorescence quantification of FADU control (grey), dermal (yellow) and oral (blue) FB secretomes (two-sided Mann–Whitney U- test, n = 6 replicates, three independent FB secretomes). f , Immunofluorescence images of FADU + dermal (left) or oral (right) FB secretomes; blue, Hoechst; orange, phalloidin; green, S1P; scale bar, 20 µm (representative images from n = 6 biologically independent wells). g , Spheroid invasion of oSCC cells treated with oral FB secretomes (grey) with 5 µM S1PR1 inhibitor (S1PR1i, ponesimod, blue) or 10 µM S1PR3 inhibitor (S1PR3i, TY-52156, yellow; Kruskal–Wallis, Dunn’s multiple comparisons, n = 5 replicates). h , Spheroid invasion of oSCC treated with oral FB secretomes (grey) with 20 µM STAT3 inhibitor (STAT3i, STX-0119, blue), 10 µM NF-κB inhibitor (NF-κBi; JSH-23, yellow) or 2 µM AKT inhibitor (AKTi; MK-2206, red; Kruskal–Wallis, Dunn’s multiple comparisons, n = 5 replicates). i , Western blot of STAT3, phospho-STAT3 Tyr705 , B-actin in oSCC (FADU) + dermal or oral FB secretomes, ±PF543, S1P rescue, S1PR1 inhibitor (ponesimod) or STAT3 inhibitor (STX-0119); numbers represent normalized pSTAT3 to total STAT3 fold change relative to oral FB secretomes, run on separate blots with independent loading controls under identical conditions (western blots representative of two independent experiments). j , oSCC basal respiration in oSCC cells treated with oral FB secretomes ± lipid stripping (Cleanascite, blue, 24 h; OCR, oxygen consumption rate; n = 24 replicates in two cell lines treated with three FB secretomes, two independent experiments, two-sided Mann–Whitney U- test). k , Mitochondrial activity in oSCC cell lines treated with oral FB secretomes (blue) measured by membrane potential-dependent immunofluorescence (two-sided Mann–Whitney U- test, n = 8 replicates, two independent cell lines). l , Mitochondrial abundance in oSCC cell lines treated with oral FB secretomes (blue) measured by immunofluorescence (two-sided Mann–Whitney U- test, n = 8 replicates, two independent cell lines). m , Basal respiration in oSCC cells treated with combinations of oral FB secretomes, 20 µM imipramine, 2 µM S1P or 20 µM STAT3i (STX-0119; n = 5 independent replicates, two-sided Mann–Whitney U- test). n , Subcutaneous tumour growth rate in NSG mice injected with oSCC UMSCC01 (grey), UMSCC01 + dermal FBs (oSCC + dermal FB, yellow) or UMSCC01 + oral FBs (oSCC + oral FB, blue; two-sided Mann–Whitney U- test, n = 7 per group). o , Ki67 proliferation quantification in oSCC (grey), oSCC + dermal FB (yellow) and oSCC + oral FBs (blue) tumours (two-sided Mann–Whitney U- test, n = 14, two measurements per tumour). p , Kaplan–Meier survival curve of oSCC (black) and oSCC + oral FBs (blue; n = 7 per group, two-sided Mantel–Cox test). Box plots show minimum to maximum values (error bars), the box indicates 25th and 75th percentiles, and the line denotes the median.

Article Snippet: STAT3 inhibitor STX-0119 (MedChemExpress, HY-103692-1ml), AKT inhibitor MK-2206 (MedChemExpress, HY-10358-1ml) and NF-κB inhibitor JSH-23 (MedChemExpress, HY-13982-1ml) were supplied at 10 mM in DMSO and diluted to 20 μM, 2 μM and 10 μM, respectively, in secretomes.

Techniques: Control, MANN-WHITNEY, Immunofluorescence, Western Blot, Stripping Membranes, Activity Assay, Membrane, Injection