Journal: Frontiers in Endocrinology

Article Title: Mechanosensitive Ion Channel Piezo1 Regulates Diet-Induced Adipose Inflammation and Systemic Insulin Resistance

doi: 10.3389/fendo.2019.00373

Figure Lengend Snippet: Piezo1 regulates adipose inflammation and lipolysis. (A,B) mRNA expression of Piezo1, IL-6, IL-1β, IL-10, TNFα, MCP-1, CD36, ATGL, HSL, LPL, and CD36 in pgWAT (A) and scWAT (B) of HFD-fed adipose-Piezo1 −/− and control mice, mRNA levels were normalized to Tbp, n = 4–7. (C–E) mRNA expression of proinflammatory and lipolytic genes in cultured primary adipocytes treated with the Piezo1 agonist Yoda1 (C) , Piezo1 antagonist GsMTx-4 (D) , or GsMTx-4 with the TLR4 inhibitor TAK-242 (E) ( n = 3–5). All data were collected from male mice. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. controls. T -test was used for (A,B,D) and one-way ANOVA was used for (C,E) . Data are from male mice and are expressed as mean ± SEM. pgWAT, perigonadal white adipose tissue; scWAT, subcutaneous white adipose tissue; HFD, high-fat diet.

Article Snippet: After 6–7 days of induction, differentiated cells were challenged with 5 or 20 μM Yoda1 (SML1558, Sigma), 1 μM TAK-242 (5.08336, Millipore) and 5 μM GsMTx-4 (STG-100, Alomone) or vehicle for 18–20 h. Cells were collected for further analysis.

Techniques: Expressing, Control, Cell Culture, Tissue, Real-time Polymerase Chain Reaction

Journal: Biochemical and biophysical research communications

Article Title: Yoda1-induced phosphorylation of Akt and ERK1/2 does not require Piezo1 activation

doi: 10.1016/j.bbrc.2018.02.058

Figure Lengend Snippet: HCAECs were pre-incubated with vehicle control or the Piezo1 channel inhibitor, GsMTx4 (10 µM), for 30 min prior to stimulation with Yoda1 (1.5 µM) for 5 min. Immunoblotting was then performed on cell lysates using antibodies against phosphorylated Akt (S473) and total Akt to assess the level of Akt activation (A) or against phosphorylated ERK1/2 (T202/Y204) and total ERK1/2 to assess the level of ERK1/2 activation (B). Representative blots from 3 independent experiments are shown. Bar graphs represents the quantification as fold change in the ratio of phosphorylated Akt to total Akt (A) or phosphorylated ERK1/2 to total ERK1/2 (B) compared to the control static condition, which is set to 1. The error bars indicate SE. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: GsMTx4 was purchased from both Tocris Bioscience and Alomone Labs (Jerusalem, Israel).

Techniques: Incubation, Control, Western Blot, Activation Assay

Journal: Stem Cell Research & Therapy

Article Title: Tensile stress promotes the chondrogenic ability of condylar cartilage stem/progenitor cells in the temporomandibular joint via the Piezo1-Ca 2+ -Prkca pathway

doi: 10.1186/s13287-025-04439-7

Figure Lengend Snippet: MA enhances the expression of chondrogenic-related genes, Piezo1, and protein kinase C alpha (Prkca) in the superficial layer of rat condylar cartilage. A The principal component analysis of transcriptome sequencing samples (the superficial layers of rat condylar cartilage after 1 week of Sham or MA operation), n = 5. B Transcriptome sequencing reveals the 15 most differentially expressed genes in the superficial layer of rat condylar cartilage between the Sham and MA groups in a heatmap, n = 5. Arrows indicate the genes of interest. C The volcano plot of the differentially expressed genes between the Sham and MA groups. D qPCR detection of chondrogenic markers, Mmp13, Piezo1, and Prkca in the superficial layer of rat condylar cartilage after 1 week of Sham or MA operation, n = 5. Data are presented as mean ± SD. ** p < 0.01, *** p < 0.001

Article Snippet: CSPCs were pretreated with 10 μM calcium chelating agent BAPTA (Abcam, USA) for 3 h before loading to block Ca 2+ influx or with 5 μM Piezo1 inhibitor GsMTx4 (TargetMol, Shanghai, China) for 3 h before loading.

Techniques: Expressing, Sequencing

Journal: Stem Cell Research & Therapy

Article Title: Tensile stress promotes the chondrogenic ability of condylar cartilage stem/progenitor cells in the temporomandibular joint via the Piezo1-Ca 2+ -Prkca pathway

doi: 10.1186/s13287-025-04439-7

Figure Lengend Snippet: Tensile loading enhances the expression of chondrogenic-related genes, Piezo1, and Prkca in rat condylar CSPCs. A The flow cytometric analysis of surface markers CD90, CD29, CD45, and CD11b/c in isolated rat condylar CSPCs at the second passage. B Representative pictures of oil red, alizarin red, and alcian blue staining after trilineage differentiation induction of rat condylar CSPCs in vitro. Scale bar: 100 μm. C , D qPCR detection (n = 6 biological replicates) and representative western blots of chondrogenic markers and Prkca in rat CSPCs after 10% tensile loading for 24 or 48 h in vitro. Full-length blots are presented in Figure S1. E , F Representative Piezo1 immunofluorescence staining images of rat condylar CSPCs after 10% tensile loading for 24 or 48 h in vitro, and the corresponding quantitative analysis, n = 3 biological replicates. Scale bar: 100 μm. Data are presented as mean ± SD. ** p < 0.01, *** p < 0.001; ns, not significant

Article Snippet: CSPCs were pretreated with 10 μM calcium chelating agent BAPTA (Abcam, USA) for 3 h before loading to block Ca 2+ influx or with 5 μM Piezo1 inhibitor GsMTx4 (TargetMol, Shanghai, China) for 3 h before loading.

Techniques: Expressing, Isolation, Staining, In Vitro, Western Blot, Immunofluorescence

Journal: Stem Cell Research & Therapy

Article Title: Tensile stress promotes the chondrogenic ability of condylar cartilage stem/progenitor cells in the temporomandibular joint via the Piezo1-Ca 2+ -Prkca pathway

doi: 10.1186/s13287-025-04439-7

Figure Lengend Snippet: Blocking Piezo1 and Ca 2+ influx reduces the chondrogenic effect of tensile loading on rat condylar CSPCs. A Representative calcium immunofluorescence staining images of rat condylar CSPCs after 10% tensile loading for 48 h with or without the pretreatment of 10 μM calcium chelator BAPTA or 5 μM Piezo1 inhibitor GsMTx4. Scale bar: 100 μm. B Western blot detection and the corresponding quantitative analysis of chondrogenic markers and Prkca in rat condylar CSPCs after 10% tensile loading for 48 h with or without the pretreatment of 10 μM calcium chelator BAPTA, n = 3 biological replicates. Full-length blots are presented in Figure S2. Data are presented as mean ± SD. * p < 0.05; ns, not significant

Article Snippet: CSPCs were pretreated with 10 μM calcium chelating agent BAPTA (Abcam, USA) for 3 h before loading to block Ca 2+ influx or with 5 μM Piezo1 inhibitor GsMTx4 (TargetMol, Shanghai, China) for 3 h before loading.

Techniques: Blocking Assay, Immunofluorescence, Staining, Western Blot

Journal: American journal of physiology. Lung cellular and molecular physiology

Article Title: Mechanical Stretch Promotes Sustained Proliferation and Inflammation in Developing Human Airway Smooth Muscle

doi: 10.1152/ajplung.00070.2025

Figure Lengend Snippet: The effect of stretch in Piezo protein expression in fASM cells. ( A , B) : fASM cells grown on Flexcell plates were exposed to 5%, 10% or 15% of static stretch for 24h. Piezo1 and Piezo2 protein expression was determined by JESS assays. The expression levels in stretched cells were normalized to that of control cells without stretch treatment. Representative JESS images are shown in ( C ). SS: static stretch. Total protein signal values (x 10 5 ) are shown at the bottom of each lane. ( D , E) : fASM cells grown on Flexcell plates were exposed to 5% or 10% of static stretch superimposed on 5% oscillatory stretch (CPAP) for 24h. The Piezo1 and Piezo2 protein expression levels in these cells were normalized to that of control cells with 5% oscillatory stretch only. Representative JESS images are shown in ( F ). Data are expressed as means ± SD (n = 6–7 cell lines, each dot represents one cell line). Open symbol: male. Solid symbol: female. All data pass Shapiro-Wilk Normality tests. For statistical analysis, paired t tests are applied to control and treatment groups. Exact P value is above the data in Fig. 1A .

Article Snippet: In some experiments, fASM cells were treated with Piezo1 antagonist GsMTx4 (1 μM, MedChemExpress, HY-P1410) or Piezo2 antagonist D-GsMTx4 (1 μM, MedChemExpress, HY-P1410B) for one hour prior to stretch and the treatments continued thereafter during the stretch protocol.

Techniques: Expressing, Control

Journal: American journal of physiology. Lung cellular and molecular physiology

Article Title: Mechanical Stretch Promotes Sustained Proliferation and Inflammation in Developing Human Airway Smooth Muscle

doi: 10.1152/ajplung.00070.2025

Figure Lengend Snippet: The long-term effect of stretch in fASM cell proliferation. fASM cells on Flexcell plates were pretreated with inhibitors to Piezo1 ( A ) or Piezo2 ( B ) for 1h and then were exposed to 5% static stretch with 5% oscillatory stretch (5% CPAP) with or without the inhibitors. Control cells were exposed to 5% oscillatory stretch only. Cells were harvested after 24h treatment and seeded into 96-well plates. After 3-day culture without stretch, cell proliferation was measured by CyQUANT assay. Open symbol: male. Solid symbol: female. All data pass Shapiro-Wilk Normality tests. For statistical analysis, Two-Way ANOVA with uncorrected Fisher’s LSD test are applied. Data are expressed as means ± SD (n = 6–7 cell lines, each dot represents one cell line). The interaction p-values are below the corresponding panels.

Article Snippet: In some experiments, fASM cells were treated with Piezo1 antagonist GsMTx4 (1 μM, MedChemExpress, HY-P1410) or Piezo2 antagonist D-GsMTx4 (1 μM, MedChemExpress, HY-P1410B) for one hour prior to stretch and the treatments continued thereafter during the stretch protocol.

Techniques: Control, CyQUANT Assay

Journal: Cancers

Article Title: Contribution of Oxidative Stress Induced by Sonodynamic Therapy to the Calcium Homeostasis Imbalance Enhances Macrophage Infiltration in Glioma Cells

doi: 10.3390/cancers14082036

Figure Lengend Snippet: Immunofluorescence of Ca 2+ under mitochondrial oxidative stress. ( A ) Immunofluorescence images showing the fluorescence intensity of Ca 2+ (Green) and MitoSOX (Red) in U251 cells under six different conditions. Groups were exposed to CON, US (40 kHz, 0.5 W/cm 2 ), SDT (10 μg/mL HMME + US), SG (500 nM GsMTx4 + SDT), SY (10 nM Yoda 1 + SDT), and SN (5 mM NAC + SDT) for 24 h. High Ca 2+ and MitoSOX immunosignal expression was observed at the intracellular and around the membrane level as well as inside the nucleus (SDT group). The prominence of the cell membrane vesicles in the SG group. ( B ) Summary data of Ca 2+ and MitoSOX expression in the six different treatment groups. In the SG group, the fluorescence intensities of Ca 2+ (green) and MitoSOX (red) were stronger than they were in the other groups. The Ca 2+ and MitoSOX fluorescence levels were analyzed using confocal analysis software (ZEN Blue Lite). Scale bar, 5 µm.

Article Snippet: It was dissolved in 10 mL sterile DMEM (Gibco, Thermo Scientific, Waltham, MA, USA) at a ratio of 1:1000 at the final concentration of 10 μg/mL and a pH of 7.4 and was liquefied at room temperature (RT) and stored in the dark at 4 °C; (2) a 1 mg amount of the Piezo1 antagonist GsMTx4 (CAS: TP1300, Topscience, Shanghai, China) was dissolved in 0.047 mL sterile double distilled water at a ratio of 1:1000 at the final concentration of 500 nM and was liquefied at RT and stored in the dark at −20 °C; (3) a 10 mg amount of the Piezo1 agonist Yoda 1 (CAS: T7506, Topscience, Shanghai, China) was dissolved in 2.815 mL sterile DMSO at a ratio of 1:1000 at the final concentration of 10 nM and was liquefied at RT and stored in the dark at −20 °C; (4) a 5 g amount of the ROS antagonist NAC (CAS: CSN16436, CSNpharm, Shanghai, China) was dissolved in 24 mL sterile double distilled water at a ratio of 1:100 at the final concentration of 5 mM, and it was liquefied at RT and stored in the dark at −20 °C; (5) a 50 μg amount of MitoSOX (CAS: M36008, Invitrogen, Waltham, MA, USA), a new type of mitochondrial fluorescent probe that can specifically target mitochondria to selectively detect superoxide in mitochondria, was also used in the present experiment.

Techniques: Immunofluorescence, Fluorescence, Expressing, Membrane, Software

Journal: Cancers

Article Title: Contribution of Oxidative Stress Induced by Sonodynamic Therapy to the Calcium Homeostasis Imbalance Enhances Macrophage Infiltration in Glioma Cells

doi: 10.3390/cancers14082036

Figure Lengend Snippet: ( A ) Immunofluorescence images of the co-expression of the Ca 2+ (green), lipid (Nile red), and Hoechst 33342 (blue) in U251 cells under six different conditions over 24 h. The data show good synchronous change trends. ( C ) Pearson correlation coefficient (PCC) and Manders overlap coefficients (MOC) ( D ) show good co-localization (r > 0.80). ( B ) Confocal microscopic images of U251 cells treated with CON, US, SDT, SG, SY, and SN for 24 h and incubated with 2 μM Fluo-4 AM for 30 min followed by immunofluorescence staining with the antibody Piezo1 (green) and Hoechst 33342 (blue). ( E ) Immunofluorescence intensities were analyzed using confocal analysis software (ZEN Blue Lite). SDT had the highest Piezo1 expression and maintained a higher intracellular calcium level. SG showed lower Piezo1 expression than the SY group but possessed a higher Ca 2+ level. ( F ) The temperature difference in the water before the ultrasonic treatment of the cells compared to the temperature after the treatment was less than 2 °C (measurement frequency, n > 5). * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: It was dissolved in 10 mL sterile DMEM (Gibco, Thermo Scientific, Waltham, MA, USA) at a ratio of 1:1000 at the final concentration of 10 μg/mL and a pH of 7.4 and was liquefied at room temperature (RT) and stored in the dark at 4 °C; (2) a 1 mg amount of the Piezo1 antagonist GsMTx4 (CAS: TP1300, Topscience, Shanghai, China) was dissolved in 0.047 mL sterile double distilled water at a ratio of 1:1000 at the final concentration of 500 nM and was liquefied at RT and stored in the dark at −20 °C; (3) a 10 mg amount of the Piezo1 agonist Yoda 1 (CAS: T7506, Topscience, Shanghai, China) was dissolved in 2.815 mL sterile DMSO at a ratio of 1:1000 at the final concentration of 10 nM and was liquefied at RT and stored in the dark at −20 °C; (4) a 5 g amount of the ROS antagonist NAC (CAS: CSN16436, CSNpharm, Shanghai, China) was dissolved in 24 mL sterile double distilled water at a ratio of 1:100 at the final concentration of 5 mM, and it was liquefied at RT and stored in the dark at −20 °C; (5) a 50 μg amount of MitoSOX (CAS: M36008, Invitrogen, Waltham, MA, USA), a new type of mitochondrial fluorescent probe that can specifically target mitochondria to selectively detect superoxide in mitochondria, was also used in the present experiment.

Techniques: Immunofluorescence, Expressing, Incubation, Staining, Software

Journal: Cancers

Article Title: Contribution of Oxidative Stress Induced by Sonodynamic Therapy to the Calcium Homeostasis Imbalance Enhances Macrophage Infiltration in Glioma Cells

doi: 10.3390/cancers14082036

Figure Lengend Snippet: The living cell system analyzed real-time dynamic changes in the Ca 2+ and lipid contents in U251 cells. ( A , C , E ) The fluorescence intensity of the intracellular Ca 2+ in the CON, US, CG, UG, and SG groups shows a downward trend, and rises and then falls to 600 fi. in the SDT group (n = 39 cells from 3 dishes) (green arrow: addition of 500 nM GsMTx4). ( B , D , F ) The fluorescence intensity of intracellular lipids in the CON, US, SDT, CG, and UG groups showed an upward trend, and a steady decline in the SG group (n = 76 cells from 3 dishes) (red arrow: addition of 500 nM GsMTx4).

Article Snippet: It was dissolved in 10 mL sterile DMEM (Gibco, Thermo Scientific, Waltham, MA, USA) at a ratio of 1:1000 at the final concentration of 10 μg/mL and a pH of 7.4 and was liquefied at room temperature (RT) and stored in the dark at 4 °C; (2) a 1 mg amount of the Piezo1 antagonist GsMTx4 (CAS: TP1300, Topscience, Shanghai, China) was dissolved in 0.047 mL sterile double distilled water at a ratio of 1:1000 at the final concentration of 500 nM and was liquefied at RT and stored in the dark at −20 °C; (3) a 10 mg amount of the Piezo1 agonist Yoda 1 (CAS: T7506, Topscience, Shanghai, China) was dissolved in 2.815 mL sterile DMSO at a ratio of 1:1000 at the final concentration of 10 nM and was liquefied at RT and stored in the dark at −20 °C; (4) a 5 g amount of the ROS antagonist NAC (CAS: CSN16436, CSNpharm, Shanghai, China) was dissolved in 24 mL sterile double distilled water at a ratio of 1:100 at the final concentration of 5 mM, and it was liquefied at RT and stored in the dark at −20 °C; (5) a 50 μg amount of MitoSOX (CAS: M36008, Invitrogen, Waltham, MA, USA), a new type of mitochondrial fluorescent probe that can specifically target mitochondria to selectively detect superoxide in mitochondria, was also used in the present experiment.

Techniques: Fluorescence

Journal: Cancers

Article Title: Contribution of Oxidative Stress Induced by Sonodynamic Therapy to the Calcium Homeostasis Imbalance Enhances Macrophage Infiltration in Glioma Cells

doi: 10.3390/cancers14082036

Figure Lengend Snippet: Schematic of SDT’s anti-glioma mechanism. The effect of ROS on glioma cells and the enhanced Ca 2+ influx induced by sonodynamic therapy. HMME was incubated into the cytoplasm with glioma cells that could be excited by ultrasound to increase the mitochondrial ROS levels. Additionally, Piezo1 was then activated and opened, resulting in the extracellular Ca 2+ flowing into the cells immediately. The reductions in the extracellular Ca 2+ promoted macrophage M1 polarization. Intracellular lipid droplets quickly formed complexes with Ca 2+ . Large amounts of ROS promote the peroxidation of lipid droplets. At the same time, mitochondrial dysfunction is accompanied by the complexation of lipid droplets with Ca 2+ , reducing the ATP produced from the cells’ lipid droplet oxidation supplies.

Article Snippet: It was dissolved in 10 mL sterile DMEM (Gibco, Thermo Scientific, Waltham, MA, USA) at a ratio of 1:1000 at the final concentration of 10 μg/mL and a pH of 7.4 and was liquefied at room temperature (RT) and stored in the dark at 4 °C; (2) a 1 mg amount of the Piezo1 antagonist GsMTx4 (CAS: TP1300, Topscience, Shanghai, China) was dissolved in 0.047 mL sterile double distilled water at a ratio of 1:1000 at the final concentration of 500 nM and was liquefied at RT and stored in the dark at −20 °C; (3) a 10 mg amount of the Piezo1 agonist Yoda 1 (CAS: T7506, Topscience, Shanghai, China) was dissolved in 2.815 mL sterile DMSO at a ratio of 1:1000 at the final concentration of 10 nM and was liquefied at RT and stored in the dark at −20 °C; (4) a 5 g amount of the ROS antagonist NAC (CAS: CSN16436, CSNpharm, Shanghai, China) was dissolved in 24 mL sterile double distilled water at a ratio of 1:100 at the final concentration of 5 mM, and it was liquefied at RT and stored in the dark at −20 °C; (5) a 50 μg amount of MitoSOX (CAS: M36008, Invitrogen, Waltham, MA, USA), a new type of mitochondrial fluorescent probe that can specifically target mitochondria to selectively detect superoxide in mitochondria, was also used in the present experiment.

Techniques: Incubation, Produced

Journal: ACS nano

Article Title: Magnetic Vortex Nanodiscs Enable Remote Magnetomechanical Neural Stimulation

doi: 10.1021/acsnano.0c00562

Figure Lengend Snippet: a, Fluorescence traces resulting from stimulation of DRG neurons incubated with 226 nm diameter MNDs (left) and 98 nm diameter MNDs with MF frequencies of 5 Hz (top) and 1 Hz (bottom). MF amplitude is sequentially increased from 7 mT to 28 mT (marked by the shaded regions). MF is applied in 4 pulses of 10 s with 30 s wait times between pulses. b, Summary of cell response rate for conditions permuted in (a). Error bars represent standard error of the mean. c, DRGs incubated with 1 μM piezo inhibitor GsMTx4 (left) and with TRPV4 antagonist HC-067047 (right) both show a decrease in activity after the first stimulation sequence. d, Confocal images of HEK-293 cells loaded with the calcium indicator Fluo-4 and transfected with the mechanosensitive TRPV4 channel labeled with mCherry. Scale bars = 100 μm. e, The response of unmodified HEK-293 cells decorated with MNDs to MF application (left), the response of HEK-293 cells expressing TRPV4 decorated with MNDs to MF application (middle), and the blocked response of HEK-293 cells decorated with MNDs, expressing TRPV4, and incubated 1 μM TRPV4 antagonist, HC-067047 (right). In experiments shown in (c) and (e), a 5 Hz, 26 mT MF is applied in 3 sequences of 10 s as shown in the shaded grey regions. The number of cells per condition is 300. Standard error is represented by shaded area in fluorescence traces.

Article Snippet: The PIEZO-specific inhibitor GsMTx4 (Smartox) was added to Tyrode’s solution at a concentration of 1 μM.

Techniques: Fluorescence, Incubation, Activity Assay, Sequencing, Transfection, Labeling, Expressing