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: 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: 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