sb202190 p38  (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: 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: This model illustrates the proposed three-tiered molecular switch that translates external microbial cues into the irreversible developmental fate of sea star metamorphosis, based on Dynamic Network Module (DNM) analysis and comprehensive pharmacological functional assays. This cascade integrates innate immune and developmental signaling pathways across three functional layers: Signal Sensing, Commitment Conversion, and Irreversible Execution. The process is initiated in the Signal Sensing layer, where the environmental cue, microbial biofilms, activates the adapter protein MyD88, which serves as an obligatory first-tier hub. MyD88 transmits signals via the JNK/p38/ERK MAPK pathway to govern the initial settlement behavior. MyD88 exhibits a concentration-dependent dual output: high-dose inhibition abolishes settlement behavior (RA-non-rescuable), while low-dose inhibition permits settlement but causes a late-stage molecular arrest (RA-non-rescuable). Following sensing, the cascade enters the Commitment Conversion layer. JNK/p38 MAPK acts as an essential hybrid adapter that converts immune signals into a Retinoic Acid (RA) hormonal commitment signal (RA-rescuable phenotype). The Amyloid Precursor Protein (APP) functions as the irrevocable commitment gateway, integrating inputs from the upstream MAPK, IKKβ/NFκB, and RA signaling axes to make the final molecular decision. APP ensures irreversibility through “signal focusing,” maintaining its signal strength during the systemic “mass shutdown” of non-essential larval programs. The process culminates in an Irreversible Execution Tier, where the robust execution of the metamorphic program relies on the multi-layered convergence of signals onto the master transcription factor, TFAP2A. The APP commitment decision is translated into transcriptional output via the release of its intracellular domain (AICD), which acts as the final dedicated execution switch by converging to TFAP2A in complex with GSK3β/Src. TFAP2A receives parallel inputs from RA (for launching the program), IKKβ/NFκB (for sustained maintenance and transcriptional output; RA non-rescuable), and ERK (a crucial early execution factor for immediate morphogenesis and physical attachment maintenance; RA non-rescuable). Finally, the RA signal induces the HSP90AA1 chaperone, establishing a positive feedback loop that maintains the structural integrity and function of critical signaling complexes (including MyD88 and APP), thereby ensuring the stability of the executed program.

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: Functional Assay, Protein-Protein interactions, Concentration Assay, Inhibition

Journal: PLOS One

Article Title: Xiao Qing Long Tang ameliorates neutrophil extracellular trap-dendritic cells-T helper 17 cell axis in Neutrophilic Asthma

doi: 10.1371/journal.pone.0336333

Figure Lengend Snippet: BMDCs were pretreated with control culture, Nets, OVA/LPS, OVA/LPS/Nets and then coculture with naïve CD4 + T lymphocytes. a. Representative flow cytometric analysis and comparisons of Th17 in each group. b Comparisons of concentrations of IL-17 in each group. c. Comparisons of concentrations of IL-6 in each group, d. Comparisons of concentrations of IL-23 in each group. e. Representative Western blot images and comparisons of p-p38/p38 MAPK, p-IKBα/IKBα, p65/p65, β-actin in each group. f. The images of immunohistochemical staining and comparisons for P-p38 MAPK, P-pIKBα, P-p65 expression in the CD11c+ positive cells of CON and OVA/LPS induced lung, (P-p38 in the lung were identified with DAPI (blue), P-p38 (red) and CD11c+ (green) by confocal microscopy, P-p65 in the lung were identified with DAPI (blue), P-p65 (red) and CD11c+ (green) by confocal microscopy, P-p65 in the lung were identified with DAPI (blue), P-pIKBα (red) and CD11c+ (green) by confocal microscopy). OVA/LPS/Nets-stimulated BMDCs were pretreated with control culture, OVA/LPS/Nets, p38 inhibitor (SB202190), OVA/LPS/Nets/SB202190, and then coculture with naïve CD4 + T lymphocytes, g. Representative Western blot images and comparisons of p-p38/p38 MAPK, p-IKBα/IKBα, p65/p65, β-actin. in each group, h. Comparisons of concentrations of IL-6 in each group, i. Comparisons of concentrations of IL-23 in each group. OVA/LPS/Nets-stimulated BMDCs were pretreated with control culture, OVA/LPS/Nets, NF-κB inhibitor (DHMEQ), OVA/LPS/Nets/DHMEQ, and then coculture with naïve CD4 + T lymphocytes, j. Representative Western blot images and comparisons of p-IKBα/IKBα, p65/p65, β-actin in each group, k. Comparisons of concentrations of IL-6 in each group, l. Comparisons of concentrations of IL-23 in each group. m. Representative flow cytometric analysis and comparisons of Th17 in CON, OLN, SB202190, OLNS group. n. Comparisons of concentrations of IL-17 in in CON, OLN, SB202190, OLNS group. o. Representative flow cytometric analysis and comparisons of Th17 in in CON, OLN, DHMEQ, OLND group. p Comparisons of concentrations of IL-17 in each group. (Data were means ± SEM (n = 3); * * P < 0.01) (CON: control group, Nets: Neutrophil extracellular traps group, OL: OVA/LPS group, OLN:OVA/LPS/Nets group, SB202190:a p38-MAPK specific inhibitor group, DHMEQ: a NF-κB-specific inhibitor group, OLNS:OVA/LPS/Nets/SB202190 group, OLND:OVA/LPS/Nets/DHMEQ group).

Article Snippet: To explore the signaling pathways involved in OVA/LPS/Nets-induced BMDCs in regulating Th17 cell differentiation, BMDCs were pre-treated with specific inhibitors: 10 μM p38-MAPK specific inhibitor SB202190(MCE, USA, HY-10295) and 5 μg/ml NF-κB-specific inhibitor DHMEQ (MCE, USA, HY-14645) 1 hour before being stimulated with OVA/LPS/Nets.

Techniques: Control, Western Blot, Immunohistochemical staining, Staining, Expressing, Confocal Microscopy

Journal: PLOS One

Article Title: Xiao Qing Long Tang ameliorates neutrophil extracellular trap-dendritic cells-T helper 17 cell axis in Neutrophilic Asthma

doi: 10.1371/journal.pone.0336333

Figure Lengend Snippet: BMDCs were pretreated with control culture (CON), OVA/LPS/Nets (OLN), XQLT, OVA/LPS/Nets/XQLT(OLNX) and then coculture with naïve CD4 + T lymphocytes. a. comparisons of concentrations of IL-6 in each group, b. comparisons of concentrations of IL-23 in each group, c. Representative flow cytometric analysis and comparisons of Th17 cells in each group, d. Comparisons of concentrations of IL-17 in each group, f. Representative Western blot images and comparisons of p-p38/p38 MAPK, p-IKBα/IKBα, p-p65/p65, β-actin in each group, f. The images of immunohistochemical staining and comparisons for P-p38 MAPK, P-pIKBα, P-p65 expression in the CD11c+ positive cells of lung, (P-p38 in the lung were identified with DAPI (blue), P-p38 (red) and CD11c+ (green) by confocal microscopy, P-p65 in the lung were identified with DAPI (blue), P-p65 (red) and CD11c+ (green) by confocal microscopy, P-p65 in the lung were identified with DAPI (blue), P-pIKBα (red) and CD11c+ (green) by confocal microscopy) (Data are means ± SEM (n = 3); ** P < 0.01) (CON: control group, XQLT: Xiao Qing Long Tang group, OLN: OVA/LPS/Nets group, OLNX:OVA/LPS/Nets/XQLT group).

Article Snippet: To explore the signaling pathways involved in OVA/LPS/Nets-induced BMDCs in regulating Th17 cell differentiation, BMDCs were pre-treated with specific inhibitors: 10 μM p38-MAPK specific inhibitor SB202190(MCE, USA, HY-10295) and 5 μg/ml NF-κB-specific inhibitor DHMEQ (MCE, USA, HY-14645) 1 hour before being stimulated with OVA/LPS/Nets.

Techniques: Control, Western Blot, Immunohistochemical staining, Staining, Expressing, Confocal Microscopy