Journal: The Journal of Clinical Investigation

Article Title: BM-derived mesenchymal stem cell microvesicles protect enteric neural precursor cells and alleviate diabetes-associated enteric neuropathy

doi: 10.1172/JCI192437

Figure Lengend Snippet: ( A ) CCK-8 assay assessing the viability of ENPCs treated with ERK inhibitor PD98059. n = 7. ( B and C ) Representative immunofluorescence images ( B ) and quantification ( C ) of EdU incorporation in ENPCs treated with ERK inhibitor PD98059. Scale bars: 20 μm. n = 6. ( D ) Flow cytometric analysis of apoptosis using annexin V/PI staining in ENPCs treated with ERK inhibitor PD98059. n = 3. ( E ) Representative immunofluorescence images of neuronal (HuC/D + and nNOS + ) and glial (GFAP + ) differentiation in ENPCs treated with ERK inhibitor PD98059. Scale bars: 100 μm. n = 3. ( F ) Western blot analysis of GRP78, p-PERK, PERK, p-EIF2A, EIF2A, ATF4, CHOP, BCL-2, and BAX proteins in ENPCs treated with ERK inhibitor PD98059. n = 3. ( G ) Western blot analysis of puromycin incorporation assessing global protein synthesis rates in ENPCs treated with ERK inhibitor PD98059. n = 3. ( H ) Representative immunofluorescence images of PROTEOSTAT-detected protein aggregation in ENPCs treated with ERK inhibitor PD98059. Scale bars: 10 μm. n = 4. Data are presented as mean ± SD. Statistical significance was determined by 1-way ANOVA with Tukey’s multiple-comparison test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, P > 0.05.

Article Snippet: For in vitro experiments, primary ENPCs were exposed with d -glucose (30 mM; PB180418 , Procell), BMSC-MVs (0.25 μg/mL), 4-PBA (5 mM), and ERK inhibitor PD98059 (10 μM; HY-12028, MedChemExpress) for 72, 24, 4, and 2 hours.

Techniques: CCK-8 Assay, Immunofluorescence, Staining, Western Blot, Comparison

Journal: Frontiers in Endocrinology

Article Title: Unveiling the TrkA-p35/CDK5 axis: a novel therapeutic target in diabetic kidney disease

doi: 10.3389/fendo.2026.1791283

Figure Lengend Snippet: TrkA inhibitor suppresses p-TrkA-mediated upregulation of ERK, EGR1, and p35 in the MPC5 cells grown under HG conditions. Representative western blots show the levels of (A) TrkA and phospho-TrkA (Tyr490), (C) ERK, phospho-ERK, and EGR1, and (E) CDK5 and p35 in the LG, HG+Vehicle (0.1% DMSO), HG + Inh-TrkA, HG + empty vector, HG + TrkA OE, and HG + TrkA OE + PD98059 groups. Representative western blots show the levels of (B) TrkA and phospho-TrkA (Tyr490), (D) ERK, phospho-ERK, and EGR1, and (F) CDK5 and p35 in the LG, HG+si-NC, HG+si-TrkA, HG + TrkA OE+Ctrl, and HG + TrkA OE + GW441756 groups. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. p < 0.05 was considered statistically significant.

Article Snippet: The ERK pathway inhibitor PD98059 (HY-12028, MedChemExpress, CA, USA).

Techniques: Western Blot, Plasmid Preparation

Journal: Frontiers in Endocrinology

Article Title: Unveiling the TrkA-p35/CDK5 axis: a novel therapeutic target in diabetic kidney disease

doi: 10.3389/fendo.2026.1791283

Figure Lengend Snippet: TrkA inhibition alleviates inflammation and injury in the HG-stimulated MPC5 cells. Representative western blots show the expression levels of (A) TNF-α and IL-1β, and (G) synaptopodin in the LG, HG+Vehicle, HG + Inh-TrkA, HG + empty vector, HG + TrkA OE, and HG + TrkA OE + PD98059 groups. Representative western blots show the expression levels of (D) TNF-α; IL-1β, and (I) synaptopodin in the LG, HG+si-NC, HG+si-TrkA, HG + TrkA OE+Ctrl, and HG + TrkA OE + GW441756 groups. Quantitative analysis shows the relative gene expression levels of (B) TNF-α; (C) IL-6; and (H) Nephrin in the LG, HG+Vehicle, LG + Man, HG + Inh-TrkA, HG + empty vector, HG + TrkA OE, and HG + TrkA OE + PD98059 groups. (E) TNF-α; (F) IL-1β; and (J) Nephrin in the LG, HG+si-NC, HG+si-TrkA, HG + TrkA OE+Ctrl, and HG + TrkA OE + GW441756 groups. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. p < 0.05 was considered statistically significant.

Article Snippet: The ERK pathway inhibitor PD98059 (HY-12028, MedChemExpress, CA, USA).

Techniques: Inhibition, Western Blot, Expressing, Plasmid Preparation, Gene Expression

Journal: Frontiers in Endocrinology

Article Title: Unveiling the TrkA-p35/CDK5 axis: a novel therapeutic target in diabetic kidney disease

doi: 10.3389/fendo.2026.1791283

Figure Lengend Snippet: TrkA inhibition alleviates apoptosis in the HG-stimulated MPC5 cells. (A) Flow cytometry plots show the analysis of the percentage of apoptotic cells in the LG, HG+Vehicle, HG + Inh-TrkA, HG + empty vector, HG + TrkA OE, and HG + TrkA OE + PD98059 groups based on Annexin V/PI double staining. PI distinguishes viable cells from apoptotic cells; Annexin V-FITC specifically identifies apoptotic cells. (B) Quantitative analysis of the percent apoptotic cells in different groups based on data shown in (A) . (C) The analysis of the percentage of apoptotic cells in the LG, HG+si-NC, HG+si-TrkA, HG + TrkA OE+Ctrl, and HG + TrkA OE + GW441756 groups. (D) Quantitative analysis of the percent apoptotic cells in different groups based on data shown in (C) . *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. p < 0.05 was considered statistically significant.

Article Snippet: The ERK pathway inhibitor PD98059 (HY-12028, MedChemExpress, CA, USA).

Techniques: Inhibition, Flow Cytometry, Plasmid Preparation, Double Staining