egfr inhibitor ag1478  (MedChemExpress)

Journal: Sexual Medicine

Article Title: Hinokiflavone alleviates high-fat diet-induced erectile dysfunction via the EGFR/PI3K/Akt/eNOS signaling pathway

doi: 10.1093/sexmed/qfaf059

Figure Lengend Snippet: HINO alleviates PA-induced endothelial dysfunction by activating the EGFR and PI3K/Akt/eNOS Signaling pathways. (A-D) Representative immunoblotting images (A) for p-PI3K, PI3K, p-Akt, Akt, p-eNOS, and eNOS after treatment with EGFR inhibitor AG1478 (10 μM, 24 h) in RCCECs. Semi-quantification of p-PI3K/PI3K (B), p-Akt/Akt (C), and p-eNOS/eNOS (D) was conducted with data presented as mean ± SD (n = 3). (E-G) Representative immunoblotting images (E) for p-Akt, Akt, p-eNOS, and eNOS after treatment with PI3K inhibitor LY294002 (10 μM, 24 h) in RCCECs. Semi-quantification of p-Akt/Akt (F) and p-eNOS/eNOS was conducted with data presented as mean ± SD (n = 3). (H-I) Representative immunoblotting images for p-eNOS and eNOS after treatment with Akt inhibitor MK-2206 (10 μM, 24 h) in RCCECs. Semi-quantification of p-eNOS/eNOS (I) was conducted with data presented as mean ± SD (n = 3). (J-K) Representative images (J) and semi-quantification (K) of nitric oxide release using DAF-FM from RCCECs treated with inhibitor AG1478, LY294002, MK-2206, and L-NAME. Scale bars, 100 μm. Data are presented as mean ± SD (n = 3).

Article Snippet: Additionally, an inhibitor working solution was formulated by dissolving powdered AG1478 (Cat# HY-13524, MCE, Shanghai, China), LY294002 (Cat# HY-10108, MCE, Shanghai, China), MK-2206 dihydrochloride (Cat# HY-10358, MCE, Shanghai, China), and L-NAME hydrochloride (Cat# HY-18729A, MCE, Shanghai, China) in DMSO.

Techniques: Protein-Protein interactions, Western Blot

Journal: bioRxiv

Article Title: CD163⁺/Dab2⁺ Macrophages Alleviate Cardiac Hypertrophy via Nrg2/ErbB4-Mediated Mitochondrial Reprogramming

doi: 10.1101/2025.05.22.655661

Figure Lengend Snippet: A, Representative flow plots of F4/80, CD11b, CD163, Dab2 and Nrg2 expression in polarized BMDMs. B, ELISA analysis of Nrg2 levels in CD163 - , CD163 + /Dab2 - , and CD163 + /Dab2 + macrophages (n=3). * P <0.05; ** P <0.01. C, Representative immunofluorescence images of phalloidin staining in NRVMs cultured with different medium and treated with vehicle or Ang II for 48 hours. Scale bar=100 μm. D, Relative cell surface area of NRVMs cultured with different medium following vehicle or Ang II treatment (n=6). * P <0.05; *** P <0.001; ns, not significant. E, RT-qPCR analysis of the mRNA levels of Anp and Myh7 in NRVMs cultured with different medium and treated with vehicle or Ang II for 48 hours (n=5). * P <0.05; ** P <0.01; *** P <0.001; ns, not significant. F, Representative immunofluorescence images of phalloidin staining in NRVMs treated with recombinant Nrg2 and inhibitor AG1478 in response to vehicle or Ang II treatment for 48 hours, respectively. Scale bar=100 μm. G, Relative cell surface area of NRVMs treated with recombinant Nrg2 and/or inhibitor AG1478 in response to vehicle or Ang II treatment for 48 hours, respectively (n=5). *** P <0.001; ns, not significant. H, RT-qPCR analysis of the mRNA levels of Anp, Bnp and Myh7 in NRVMs treated with recombinant Nrg2 and and inhibitor AG1478 in response to vehicle or Ang II treatment for 48 hours, respectively (n=3). * P <0.05; ** P <0.01; *** P <0.001. I, Protein expression of Anp and Myh7 in NRVMs treated with recombinant Nrg2 and inhibitor AG1478 in response to vehicle or Ang II treatment for 48 hours, respectively

Article Snippet: For the activation and blockade of the Nrg2 and ErbB4 pathway, 1 week after TAC surgery, mice were randomized and continuously treated with either vehicle or recombinant Nrg2 (produced by our lab) at a dosage of 500 μg/kg/d and inhibitor AG1478 (HY-13524, MCE) at a dosage of 10 mg/kg/d for 7 days.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Cell Culture, Quantitative RT-PCR, Recombinant

Journal: bioRxiv

Article Title: CD163⁺/Dab2⁺ Macrophages Alleviate Cardiac Hypertrophy via Nrg2/ErbB4-Mediated Mitochondrial Reprogramming

doi: 10.1101/2025.05.22.655661

Figure Lengend Snippet: A, Schematic diagram illustrating the experimental design involving the injection of recombinant Nrg2 and inhibitor AG1478 into a mouse model of cardiac hypertrophy. Wildtype mice at 6 weeks of age were subjected to transverse aortic constriction (TAC) or Sham surgery. After 1 week, recombinant Nrg2 or Nrg2 and inhibitor AG1478 were continuously administered into the mice for 7 days. These mice groups were denoted as Sham, TAC, TAC+Nrg2, and TAC+Nrg2+AG1478. B, Representative images of hearts at 8 weeks post-surgery from mice subjected to Sham or TAC surgery and injected with recombinant Nrg2 or Nrg2 and inhibitor AG1478. C, Heart weight/body weight (HW/BW) ratios at 8 weeks post-surgery for mice subjected to Sham or TAC surgery and injected with recombinant Nrg2 or Nrg2 and inhibitor AG1478 (Sham, n=8; TAC, n=7; TAC+Nrg2, n=8; TAC+Nrg2+AG1478, n=8). ** P <0.01; *** P <0.001; ns, not significant. D and E, M-mode echocardiography images (D) and quantified parameters (E) of the left ventricular (LV) chamber at 8 weeks post-surgery in mice subjected to Sham or TAC surgery and injected with recombinant Nrg2 or Nrg2 and inhibitor AG1478 (Sham, n=8; TAC, n=7; TAC+Nrg2, n=8; TAC+Nrg2+AG1478, n=8). ** P <0.01; *** P <0.001; ns, not significant. EF, ejection fraction; FS, fractional shortening; CO, cardiac output. F, Hematoxylin-eosin (H&E) (scale bar=1 mm) and Wheat germ agglutinin (WGA) staining (scale bar=50 μm) showing morphology and cell boundaries of heart sections 8 weeks after TAC with indicated treatment regimens. G, Quantification of the cardiomyocyte cross-sectional areas of the indicated groups (n=6 mice per group). *** P <0.001; ns, not significant. H, The mRNA levels of hypertrophic marker genes Anp, Bnp, Myh7 and Acta1 in hearts of the indicated groups (n=6 mice per group). * P <0.05; *** P <0.001; ns, not significant. I, The mRNA levels of fibrosis marker genes Col1a1, Col3a1, Ctgf and MMP9 in hearts of the indicated groups (n=6 mice per group). * P <0.05; ** P <0.01; *** P <0.001; ns, not significant. J, Masson’s trichrome staining showing fibrosis in heart sections 8 weeks after TAC with indicated treatment regimens. Scale bar=50 μm. K, Quantification of cardiac fibrosis in tissue shown in panel J (n=6 mice per group). * P <0.05; *** P <0.001.

Article Snippet: For the activation and blockade of the Nrg2 and ErbB4 pathway, 1 week after TAC surgery, mice were randomized and continuously treated with either vehicle or recombinant Nrg2 (produced by our lab) at a dosage of 500 μg/kg/d and inhibitor AG1478 (HY-13524, MCE) at a dosage of 10 mg/kg/d for 7 days.

Techniques: Injection, Recombinant, Staining, Marker