e2920 ikk16 mce  (MedChemExpress)

Journal: bioRxiv

Article Title: An APP-centered molecular gateway integrates innate immunity and retinoic acid signaling to drive irreversible metamorphic commitment

doi: 10.64898/2026.01.22.700939

Figure Lengend Snippet: (A) Settlement rate of inhibitor-treated larvae. The box plots with superimposed jitter plots display the larval settlement rate under various inhibitor treatments. The biofilm stimulus condition is used as the positive control. The concentration of each inhibitor is indicated on the horizontal axis. The data represent the settlement rate of larvae remaining attached out of 10 larvae across six independent biological replicates (total n = 60). Statistical significance among treatment groups was assessed using one-way ANOVA followed by Tukey’s HSD post hoc test, with grouping letters indicating significant differences ( p < 0.05); treatments sharing a letter are not significantly different. (B) Quantitative assessment of the functional hierarchy. The box plots with superimposed jitter plots show the Metamorphic Progression Scores (MPS) for larvae treated with various pharmacological inhibitors with or without all-trans retinoic acid (RA). The MPS was calculated based on the metamorphic stage reached by the larvae in the identical assays used for the settlement rate analysis in (A). The concentration of each inhibitor is indicated on the horizontal axis. The MPS represents the average metamorphic stage reached (0 = brachiolaria; 1 = early; 2 = middle; 3 = late; 4 = pre-juvenile; 5 = juvenile). Statistical significance among the treatment groups was assessed using one-way ANOVA followed by Tukey’s HSD post hoc test ( *p < 0.05; n.s., not significant). A significant RA-dependent rescue condition (a statistically significant increase in MPS compared with the inhibitor-alone condition) is highlighted in grey, establishing the functional hierarchy of the pathways relative to the RA commitment signal. (C) Representative image illustrating pathway functional hierarchy. Images show representative larval morphology under the control, inhibitor-only, and inhibitor + RA conditions. These images specifically represent the high-concentration inhibitor treatments (MyD88 inhibitor: 50 µM; MAPK inhibitors: 10 µM; IKKβ and HSP90AA1 inhibitors: 1 µM). MyD88 inhibition completely blocks the behavioral decision of settlement. JNK and p38 inhibition caused a distinct early-stage arrest (low MPS), and the effects of their inhibition were significantly rescued by RA co-treatment. In contrast, ERK inhibition arrested metamorphosis at the middle stage, and this block was not rescued by exogenous RA. Similarly, IKKβ and HSP90AA1 inhibition arrested metamorphosis at later stages, and this block was not rescued by exogenous RA, functionally placing all three pathways (ERK, IKKβ, and HSP90AA1) downstream of the RA commitment signal. Scale bar: 200 µm. Inhibitors used: T6167923 (MyD88 inhibitor), IKK-16 (IKKβ inhibitor), U0126 (ERK inhibitor), SP600125 (JNK inhibitor), SB202190 (p38 inhibitor), and Luminespib (HSP90AA1 inhibitor).

Article Snippet: The inhibitors were dissolved in DMSO and applied at the indicated concentrations: the MyD88 inhibitor T6167923 (5 or 50 μM; MedChemExpress), the IKKβ inhibitor IKK-16 (0.1 or 1 μM; MedChemExpress), and MAPK inhibitors SP600125 (JNK), SB202190 (p38), and U0126 (ERK) (1 or 10 μM; MedChemExpress or FUJIFILM Wako Pure Chemical Corporation), and the HSP90AA1 inhibitors luminespib (0.1 or 1 μM; Chemscene).

Techniques: Positive Control, Concentration Assay, Functional Assay, Control, Inhibition, Blocking Assay

Journal: mSphere

Article Title: The Pseudomonas aeruginosa effector protein TesG regulates alternative activation of macrophages through NLRC5

doi: 10.1128/msphere.00681-25

Figure Lengend Snippet: TesG regulates the polarization of macrophages through known functions of NLRC5. ( A ) iBMDMs were treated overnight with either TesG protein (10 µg/mL) alone or in combination with NLRC5 pathway inhibitors (5 µM Ac-YVAD-cmk, caspase-1 inhibitor; 1 µM peficitinib, JAK inhibitor; and 0.1 µM IKK16, IKK-2 inhibitor), and macrophage polarization was analyzed by flow cytometry. Data are presented as mean ± SEM of three to four biological replicates, with statistical significance determined by one-way ANOVA with Tukey’s multiple comparison test: * P < 0.05; ** P < 0.01; *** P < 0.001; and ns, not significant.

Article Snippet: Adherent cells in 12-well plates were incubated overnight with TesG (10 μg/mL) together with DMSO (vehicle control) or in combination with one of the following inhibitors: Ac-YVAD-cmk (5 μM), peficitinib (1 μM), or IKK16 (0.1 μM) (all from MedChemExpress).

Techniques: Flow Cytometry, Comparison

Journal: Science Advances

Article Title: Dual mechanism of inflammation sensing by the hematopoietic progenitor genome

doi: 10.1126/sciadv.adv3169

Figure Lengend Snippet: ( A ) ATAC-seq profiles depicting chromatin accessibility of the top 10 genes with inflammation-induced (left) and inflammation-independent PU.1 occupancy (right). ATAC-seq profiles in hi-77 −/− cells without inflammation were mined from published data ( GSE201968 ) at loci harboring PU.1 occupancy (CUT&Tag replicate 2) in vehicle-treated (−) and inflammation-treated (+) hi-77 −/− cells. ( B ) Comparison of average ATAC-seq signals from four biological replicates between genes with inflammation-induced PU.1 occupancy and genes with inflammation-independent PU.1 occupancy in (A). ( C ) Dose-response curve of three representative genes from inflammation-induced PU.1 occupancy and inflammation-independent PU.1 occupancy to IKK inhibitor BMS-345541. hi-77 −/− progenitors were pretreated with increasing concentrations of BMS-345541 for 1 hour and treated with both IFN-γ (1 ng/ml) and Pam 3 CSK 4 (100 ng/ml) for 4 hours ( n = 6 biological replicates). hi-77 +/+ treated with or without both agents served as negative controls. Median inhibitory concentration (IC 50 ) for each gene was calculated using nonlinear regression. Cd69 IC 50 was uncalculated. ( D ) The IC 50 of genes with inflammation-induced PU.1 occupancy was compared to those of genes with inflammation-independent occupancy. Individual values in (B) and (D) and means ± SEM in (B) to (D) were shown. Unpaired t test in (B) and (D) and multiple unpaired t test in (C). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: IKKβ inhibitors are from Selleckchem [BMS-345541 (S8044)] or a gift from S. Miyamoto (UW-Madison) [IKK16 (S2882)].

Techniques: Comparison, Concentration Assay

Journal: Science Advances

Article Title: Dual mechanism of inflammation sensing by the hematopoietic progenitor genome

doi: 10.1126/sciadv.adv3169

Figure Lengend Snippet: The hematopoietic progenitor genome uses a dual PU.1-dependent mechanism to sense and respond to inflammation. A gene cohort has inaccessible chromatin in the steady state, and inflammation increases chromatin accessibility and occupancy of the hematopoietic transcription factors GATA2 and PU.1 to activate transcription via an IKKβ-dependent mechanism. At another inflammation-activated gene cohort, GATA2 and PU.1 occupancy precede inflammation, and this mechanism is not compromised by IKKβ inhibition. RUNX1 co-occupies chromatin with GATA2 and PU.1, yet GATA2 and RUNX1 differentially control inflammation-activated transcription, and transcriptional responses involving distinct TLR signaling pathways activated by unique pathogen-associated molecular patterns .

Article Snippet: IKKβ inhibitors are from Selleckchem [BMS-345541 (S8044)] or a gift from S. Miyamoto (UW-Madison) [IKK16 (S2882)].

Techniques: Inhibition, Control, Protein-Protein interactions