hy p1410  (MedChemExpress)

Journal: Journal of Translational Medicine

Article Title: Piezo1 mediates hypoxia-induced endometriosis fibrosis via the mtDNA -dependent cGAS-STING pathway

doi: 10.1186/s12967-026-08255-2

Figure Lengend Snippet: Piezo1 expression was upregulated in ectopic lesions from patients with endometriosis. (A , B) Masson’s trichrome and Sirius Red staining show collagen deposition (indicative of fibrosis) in normal endometrial (NM), eutopic endometrial (Eu), and ectopic endometrial (Ec) (Scale bar: 100 μm). (C , D) Immunofluorescence analysis of Piezo1, Collagen I (a major ECM component, indicator of collagen deposition), and α-SMA (a myofibroblast activation marker) expression in NM, Eu, and Ec tissues. Representative images are shown (Scale bar: 100 μm). (E , F) Immunohistochemical (IHC) analysis and quantification of HIF-1α and Piezo1 protein expression in NM, Eu, and Ec tissues (Scale bar: 100 μm). (G , H) Western blotting analysis and quantification of HIF-1α, Piezo1, cGAS, and STING protein levels in NM, Eu, and Ec tissues ( n = 9). *p < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: At 60-70% confluency, cells were treated for 24 h with the specified compounds: the Piezo1 agonist Yoda1 (5 μM; MCE, HY-18723); the Piezo1 inhibitor GsMTx4 (5 μM; MCE, HY-P1410); Ethidium bromide (0.2 μg/ml; Macklin, 1239-45-8) to deplete mitochondrial DNA; the calcium chelator BAPTA-AM (4 μM; MCE, HY-100168); the cGAS inhibitor RU.521(2.5μM, MCE, HY-114180 ); the STING inhibitor C-176 (2.5 μM; MCE, HY-112906); the anti-IL6 antibody (2.5 μg/ml, Sigma-Aldrich, I7901); or the ROS inhibitor NAC (2.0mM, MCE, HY-B0215).

Techniques: Expressing, Staining, Immunofluorescence, Activation Assay, Marker, Immunohistochemical staining, Western Blot

Journal: Journal of Translational Medicine

Article Title: Piezo1 mediates hypoxia-induced endometriosis fibrosis via the mtDNA -dependent cGAS-STING pathway

doi: 10.1186/s12967-026-08255-2

Figure Lengend Snippet: Hypoxia (24 H) induces Piezo1 upregulation and fibrotic transition in HESCs. (A , B , C) The protein and mRNA expression levels of HIF-1α and Piezo1 in HESCs were analyzed by Western blotting and qRT-PCR, following exposure to hypoxia for 0,6, 12, 24, 36, 48 h, and were subsequently quantified. (D , E) The protein levels of the fibrotic markers α-SMA and Collagen I were determined by Western blotting analysis in HESCs following exposure to hypoxia for 0, 6, 12, 24, 36, 48 h. (F , G) Immunofluorescence was employed to compare α-SMA expression in HESCs subjected to different periods of hypoxic culture (Scale bar: 100 μm). (H , I) Flow cytometric analysis was used to determine the apoptosis levels in HESCs under varying hypoxic conditions. (J , K) EdU assays evaluated cell proliferation capacity under different hypoxia conditions (Scale bar: 100 μm). (L) Schematic representation of the human Piezo1 promoter, highlighting the predicted hypoxia response element (HRE) cluster (positions -220 to -180 bp) and the regions amplified by ChIP-qPCR. (M) ChIP-qPCR analysis of HIF-1α binding to the Piezo1 promoter in HESCs exposed to normoxia or hypoxia (1% O₂, 24 h). Enrichment was calculated as percentage of input (%Input). * P < 0.05 ** P < 0.01 *** P < 0.001 **** P < 0.0001

Article Snippet: At 60-70% confluency, cells were treated for 24 h with the specified compounds: the Piezo1 agonist Yoda1 (5 μM; MCE, HY-18723); the Piezo1 inhibitor GsMTx4 (5 μM; MCE, HY-P1410); Ethidium bromide (0.2 μg/ml; Macklin, 1239-45-8) to deplete mitochondrial DNA; the calcium chelator BAPTA-AM (4 μM; MCE, HY-100168); the cGAS inhibitor RU.521(2.5μM, MCE, HY-114180 ); the STING inhibitor C-176 (2.5 μM; MCE, HY-112906); the anti-IL6 antibody (2.5 μg/ml, Sigma-Aldrich, I7901); or the ROS inhibitor NAC (2.0mM, MCE, HY-B0215).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunofluorescence, Amplification, ChIP-qPCR, Binding Assay

Journal: Journal of Translational Medicine

Article Title: Piezo1 mediates hypoxia-induced endometriosis fibrosis via the mtDNA -dependent cGAS-STING pathway

doi: 10.1186/s12967-026-08255-2

Figure Lengend Snippet: Hypoxia promotes endometrial fibrosis by upregulating Piezo1. (A , B) HESCs were transfected with lentiviral vectors encoding either a non-targeting control shRNA (sh-NC) or one of two Piezo1-targeting shRNAs (sh-Piezo1-1 and sh-Piezo1-2). Subsequently, Piezo1 knockdown efficiency was analyzed by Western blotting and qRT‑PCR. (C-E) The expression levels of HIF-1α and Piezo1 in HESCs from the NC, Hypoxia, Hypoxia-sh Piezo1, and Hypoxia-Yoda1 groups were quantified using Western blotting and qRT-PCR, followed by quantitative analysis. (F , G) Western blotting was performed to detect and quantify the protein expression of fibrosis markers α-SMA and Collagen I in HESCs from the NC, Hypoxia, Hypoxia-sh Piezo1, and Hypoxia-Yoda1 groups. (H , I) Immunofluorescence staining was performed to detect α-SMA expression levels in HESCs from the NC group, hypoxia group, Hypoxia-sh Piezo1 group, and Hypoxia-Yoda1 group. Representative images were captured and subjected to quantitative analysis (Scale bar: 100 μm). (J , K) The expression of cGAS and STING was assessed by immunohistochemistry in normal endometrial tissue ( n = 9), eutopic endometrial and ectopic endometrial tissue ( n = 9). Representative images were acquired and subjected to quantitative analysis (Scale bar: 100 μm). (L-N) The expression levels of cGAS and STING in HESCs were quantified by Western blotting and qRT-PCR across the following experimental groups: NC, Hypoxia, Hypoxia-shPiezo1, and Hypoxia-Yoda1, along with their respective quantification. (O , P) Western blotting was performed to analyze TBK1 and IRF3 phosphorylation in HESCs across the following groups: NC, Hypoxia, Hypoxia-shPiezo1, and Hypoxia-Yoda1. The ratios of phosphorylated to total protein (p-TBK1/TBK1 and p-IRF3/IRF3) were calculated as indicators of pathway activation. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001

Article Snippet: At 60-70% confluency, cells were treated for 24 h with the specified compounds: the Piezo1 agonist Yoda1 (5 μM; MCE, HY-18723); the Piezo1 inhibitor GsMTx4 (5 μM; MCE, HY-P1410); Ethidium bromide (0.2 μg/ml; Macklin, 1239-45-8) to deplete mitochondrial DNA; the calcium chelator BAPTA-AM (4 μM; MCE, HY-100168); the cGAS inhibitor RU.521(2.5μM, MCE, HY-114180 ); the STING inhibitor C-176 (2.5 μM; MCE, HY-112906); the anti-IL6 antibody (2.5 μg/ml, Sigma-Aldrich, I7901); or the ROS inhibitor NAC (2.0mM, MCE, HY-B0215).

Techniques: Transfection, Control, shRNA, Knockdown, Western Blot, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Immunohistochemistry, Phospho-proteomics, Activation Assay

Journal: Journal of Translational Medicine

Article Title: Piezo1 mediates hypoxia-induced endometriosis fibrosis via the mtDNA -dependent cGAS-STING pathway

doi: 10.1186/s12967-026-08255-2

Figure Lengend Snippet: Piezo1-mediated Ca²⁺ influx promotes endometrial fibrosis via activation of the cGAS-STING pathway. (A) Immunofluorescence images showing cytosolic and mitochondrial Ca²⁺ levels in HESCs under the indicated conditions: Normoxic control (NC), Hypoxia, Hypoxia-GsMTx4, and Hypoxia-BAPTA AM (Scale bar: 100 μm). (B) Immunofluorescence analysis of mitochondrial morphology in HESCs from the NC, Hypoxia, and Hypoxia with GsMTx4 groups (Scale bar: 20 μm). (C) qRT-PCR analysis quantifying the levels of cytoplasmic mtDNA in NC and Hypoxic HESCs. (D) Representative immunofluorescence images of HESCs co-immunostained for the mitochondrial marker TOM20 and dsDNA, showing mtDNA leakage in the NC, Hypoxia, and Hypoxia-GsMTx4 groups (Scale bar: 20 μm). (E) Immunofluorescence staining analysis of the mitochondrial ROS levels in HESC cell (Scale bar: 20 μm). (F , G) Western blotting and qRT-PCR analysis of HIF-1α, Piezo1, cGAS and STING expression in HESCs Quantitative data are presented as mean ± SD. (H , I) Representative immunofluorescence images and quantification of the fibrosis marker α-SMA in HESCs (Scale bar: 100 μm). *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001

Article Snippet: At 60-70% confluency, cells were treated for 24 h with the specified compounds: the Piezo1 agonist Yoda1 (5 μM; MCE, HY-18723); the Piezo1 inhibitor GsMTx4 (5 μM; MCE, HY-P1410); Ethidium bromide (0.2 μg/ml; Macklin, 1239-45-8) to deplete mitochondrial DNA; the calcium chelator BAPTA-AM (4 μM; MCE, HY-100168); the cGAS inhibitor RU.521(2.5μM, MCE, HY-114180 ); the STING inhibitor C-176 (2.5 μM; MCE, HY-112906); the anti-IL6 antibody (2.5 μg/ml, Sigma-Aldrich, I7901); or the ROS inhibitor NAC (2.0mM, MCE, HY-B0215).

Techniques: Activation Assay, Immunofluorescence, Control, Quantitative RT-PCR, Marker, Staining, Western Blot, Expressing

Journal: Chinese Journal of Traumatology

Article Title: Piezo1 mediates electrotaxis of alveolar epithelial cells via calcium-dependent PI3K/Akt signaling

doi: 10.1016/j.cjtee.2026.01.004

Figure Lengend Snippet: Appendix 4 . Representative time-lapse video of cells under EF stimulation with GSMTx4 treatment.Multimedia component 4

Article Snippet: The Piezo1-specific inhibitor GSMTx4 (MedChemExpress HY-P1410) was used at a final concentration of 5 μM as indicated.

Techniques:

Journal: Chinese Journal of Traumatology

Article Title: Piezo1 mediates electrotaxis of alveolar epithelial cells via calcium-dependent PI3K/Akt signaling

doi: 10.1016/j.cjtee.2026.01.004

Figure Lengend Snippet: EF stimulation promotes A549 cell electrotaxis and upregulates Piezo1 expression. (A) Time-lapse images, migration trajectory plots, and quantitative analysis of A549 cells under no EF and EF (100 mV/mm) conditions. Red arrows indicate migration paths; yellow arrowheads indicate cathodal direction. Scale bar = 20 μm. (B) Quantification of Piezo1 mRNA expression by qPCR. (C) Western blot and quantification of Piezo1 protein levels. (D) Time-lapse images, migration trajectory plots, and quantitative analysis of A549 cells with or without GSMTx4 treatment under EF stimulation. ∗ p < 0.05, ∗∗∗∗ p < 0.001. EF: electric field; qPCR: quantitative polymerase chain reaction.

Article Snippet: The Piezo1-specific inhibitor GSMTx4 (MedChemExpress HY-P1410) was used at a final concentration of 5 μM as indicated.

Techniques: Expressing, Migration, Western Blot, Real-time Polymerase Chain Reaction

Journal: Chinese Journal of Traumatology

Article Title: Piezo1 mediates electrotaxis of alveolar epithelial cells via calcium-dependent PI3K/Akt signaling

doi: 10.1016/j.cjtee.2026.01.004

Figure Lengend Snippet: Inhibition of Piezo1 suppresses EF-induced calcium influx and PI3K/Akt pathway activation in A549 cells. (A) Representative images of intracellular calcium levels in A549 cells with or without GSMTx4 after EF stimulation (100 mV/mm). Scale bar = 20 μm. (B) Quantification of intracellular calcium levels. (C) Western blot and quantification of p-PI3K and p-Akt protein levels. (D) Western blot and quantification of p-PI3K and p-Akt protein levels in A549 cells pretreated with the calcium chelator BAPTA-AM under EF stimulation. EF: electric field; PI3K: phosphatidylinositol 3-kinase; Akt: protein kinase B. ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: The Piezo1-specific inhibitor GSMTx4 (MedChemExpress HY-P1410) was used at a final concentration of 5 μM as indicated.

Techniques: Inhibition, Activation Assay, Western Blot