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Novus Biologicals piezo1 mechanosensitive ion channels

Heightened <t>piezo1</t> expression around the stiff implant. A) Representative images showing Piezo1 expression around the flexible (left) and stiff (right) implant site (*). B) Quantification of normalized intensity as a function of distance from the implant (μm). C) Normalized intensity averaged across the first 50 μm from the implant site. 19 images for the flexible and 22 images for the stiff group were analyzed. D) Representative images showing Piezo1 (cyan), Iba-1 (green), GFAP (red), and merged expression around a stiff implant, with the zoomed inset with DAPI (blue) overlay. E) Correlation of total piezo1 expression with total Iba-1 expression per image for flexible (left, blue) and stiff (right, red) groups. The slope is significant for the stiff group, indicating a strong correlation between Piezo1 expression and Iba-1 intensity. F) Correlation of total piezo1 expression with total GFAP expression per image for flexible (left, blue) and stiff (right, red). 4 animals were implanted in each group. C) Two-way ANOVA with Sidak’s multiple comparison test was used, E-F) Data was normalized per image and the mean set to 0. A simple linear regression was performed, dashed lines indicate 95 % confidence intervals, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. The scale bar is 100 μm in A) and D) and 50 μm in the zoomed inset in D). C) Data are mean ± SEM. E-F) Data is sum.

Piezo1 Mechanosensitive Ion Channels, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 95/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 95 stars, based on 32 article reviews

piezo1 mechanosensitive ion channels - by Bioz Stars, 2026-05

95/100 stars

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1) Product Images from "A comparative study assessing neural recording quality and inflammatory tissue response between stiff and flexible microelectrode arrays"

Article Title: A comparative study assessing neural recording quality and inflammatory tissue response between stiff and flexible microelectrode arrays

Journal: Biomaterials

doi: 10.1016/j.biomaterials.2025.123929

Heightened piezo1 expression around the stiff implant. A) Representative images showing Piezo1 expression around the flexible (left) and stiff (right) implant site (*). B) Quantification of normalized intensity as a function of distance from the implant (μm). C) Normalized intensity averaged across the first 50 μm from the implant site. 19 images for the flexible and 22 images for the stiff group were analyzed. D) Representative images showing Piezo1 (cyan), Iba-1 (green), GFAP (red), and merged expression around a stiff implant, with the zoomed inset with DAPI (blue) overlay. E) Correlation of total piezo1 expression with total Iba-1 expression per image for flexible (left, blue) and stiff (right, red) groups. The slope is significant for the stiff group, indicating a strong correlation between Piezo1 expression and Iba-1 intensity. F) Correlation of total piezo1 expression with total GFAP expression per image for flexible (left, blue) and stiff (right, red). 4 animals were implanted in each group. C) Two-way ANOVA with Sidak’s multiple comparison test was used, E-F) Data was normalized per image and the mean set to 0. A simple linear regression was performed, dashed lines indicate 95 % confidence intervals, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. The scale bar is 100 μm in A) and D) and 50 μm in the zoomed inset in D). C) Data are mean ± SEM. E-F) Data is sum.

Figure Legend Snippet: Heightened piezo1 expression around the stiff implant. A) Representative images showing Piezo1 expression around the flexible (left) and stiff (right) implant site (*). B) Quantification of normalized intensity as a function of distance from the implant (μm). C) Normalized intensity averaged across the first 50 μm from the implant site. 19 images for the flexible and 22 images for the stiff group were analyzed. D) Representative images showing Piezo1 (cyan), Iba-1 (green), GFAP (red), and merged expression around a stiff implant, with the zoomed inset with DAPI (blue) overlay. E) Correlation of total piezo1 expression with total Iba-1 expression per image for flexible (left, blue) and stiff (right, red) groups. The slope is significant for the stiff group, indicating a strong correlation between Piezo1 expression and Iba-1 intensity. F) Correlation of total piezo1 expression with total GFAP expression per image for flexible (left, blue) and stiff (right, red). 4 animals were implanted in each group. C) Two-way ANOVA with Sidak’s multiple comparison test was used, E-F) Data was normalized per image and the mean set to 0. A simple linear regression was performed, dashed lines indicate 95 % confidence intervals, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. The scale bar is 100 μm in A) and D) and 50 μm in the zoomed inset in D). C) Data are mean ± SEM. E-F) Data is sum.

Techniques Used: Expressing, Comparison

Cell-specific intensity analysis. A) Microglia mask of a sample image using Iba-1. Black portions represent the extracted cell structures. This mask was used to identify the sample microglia shown in B). B) Piezo1 (blue) stained image showing activated microglia expressing Piezo1. C) Average within-microglia Piezo1 intensity with distance from the probe center. D) Average within-microglia Iba-1 intensity with distance from the probe center. E) Astrocyte mask of the above image using GFAP. Black portions represent the extracted cell structures. This mask was used to identify the sample astrocyte processes shown in F). F) Piezo1 (blue) stained image showing astrocyte processes expressing Piezo1. G) Average within-astrocyte Piezo1 intensity with distance from the probe center. H) Average within-astrocyte GFAP intensity with distance from the probe center. For all microglia and astrocyte analysis, the increase in intensity with proximity to the injury is highly significant for both probe substrates, and the cells surrounding stiff probes have higher intensity expression than the flexible. I) Piezo1 (red) stained image overlayed with NeuN ROIs produced by Cellpose. J) Piezo1 (red) stained image merged with NeuN (green). K) Average within-neuron Piezo1 intensity with distance from the probe center. Sample DAPI (white) stained image for a L) stiff and M) flexible probe implant. N) Average DAPI circularity index for all images analyzed above. Bar plot is the same data, but binned into two large bins rather than 10 evenly spaced bins. For A) and E), DAPI is overlayed on the mask. DAPI that passes through masked cell structures is seen as red, while DAPI covered by the mask is seen as yellow. Scale bar is 100 μm and 50 μm in zoomed insets in M&N. * Indicates implantation site. For comparisons between stiff and flexible, a two-way ANOVA with Sidak’s multiple comparison test was used. For correlation of intensity with distance from the probe, a Spearman’s r correlation test was used due to the non-normality of many of the datasets, **p < 0.01, ***p < 0.001, ****p < 0.0001. All data is mean ± SEM.

Figure Legend Snippet: Cell-specific intensity analysis. A) Microglia mask of a sample image using Iba-1. Black portions represent the extracted cell structures. This mask was used to identify the sample microglia shown in B). B) Piezo1 (blue) stained image showing activated microglia expressing Piezo1. C) Average within-microglia Piezo1 intensity with distance from the probe center. D) Average within-microglia Iba-1 intensity with distance from the probe center. E) Astrocyte mask of the above image using GFAP. Black portions represent the extracted cell structures. This mask was used to identify the sample astrocyte processes shown in F). F) Piezo1 (blue) stained image showing astrocyte processes expressing Piezo1. G) Average within-astrocyte Piezo1 intensity with distance from the probe center. H) Average within-astrocyte GFAP intensity with distance from the probe center. For all microglia and astrocyte analysis, the increase in intensity with proximity to the injury is highly significant for both probe substrates, and the cells surrounding stiff probes have higher intensity expression than the flexible. I) Piezo1 (red) stained image overlayed with NeuN ROIs produced by Cellpose. J) Piezo1 (red) stained image merged with NeuN (green). K) Average within-neuron Piezo1 intensity with distance from the probe center. Sample DAPI (white) stained image for a L) stiff and M) flexible probe implant. N) Average DAPI circularity index for all images analyzed above. Bar plot is the same data, but binned into two large bins rather than 10 evenly spaced bins. For A) and E), DAPI is overlayed on the mask. DAPI that passes through masked cell structures is seen as red, while DAPI covered by the mask is seen as yellow. Scale bar is 100 μm and 50 μm in zoomed insets in M&N. * Indicates implantation site. For comparisons between stiff and flexible, a two-way ANOVA with Sidak’s multiple comparison test was used. For correlation of intensity with distance from the probe, a Spearman’s r correlation test was used due to the non-normality of many of the datasets, **p < 0.01, ***p < 0.001, ****p < 0.0001. All data is mean ± SEM.

Techniques Used: Staining, Expressing, Produced, Comparison

Image Search Results

OBNC microspheres activate integrin receptors and mechanosensitive calcium channels. A) Mechanistic diagram of integrin activation verified using fluorophores. B-C) Fluorescence microscopy images of MSCs loaded on HAMA or OBNC hydrogel. D) Fluorescence intensity of single cell in each group was quantified. E) Fluorescence microscopy of MSCs loaded on OBNC hydrogel after different treatments. F) Fluorescence intensity in the whole field of view for each group. G) Fluorescence intensity in the single cell for each group. H) Schematic representation of patch clamp experiments. I) Electrical signals generated by MSCs in response to mechanical stimulation. J) Statistical analysis of poking currents (n = 6). K) The concentration of calcium ions in stem cells of different groups as detected by flow cytometry (siRNA1: targeting the TRPM4 gene, siRNA2: targeting the PIEZO1 gene). L) Quantitative analysis of flow cytometric results (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).

Journal: Bioactive Materials

Article Title: Mechanically sensitized hydrogel microspheres trigger membrane receptor switch for cartilage repair

doi: 10.1016/j.bioactmat.2026.03.017

Figure Lengend Snippet: OBNC microspheres activate integrin receptors and mechanosensitive calcium channels. A) Mechanistic diagram of integrin activation verified using fluorophores. B-C) Fluorescence microscopy images of MSCs loaded on HAMA or OBNC hydrogel. D) Fluorescence intensity of single cell in each group was quantified. E) Fluorescence microscopy of MSCs loaded on OBNC hydrogel after different treatments. F) Fluorescence intensity in the whole field of view for each group. G) Fluorescence intensity in the single cell for each group. H) Schematic representation of patch clamp experiments. I) Electrical signals generated by MSCs in response to mechanical stimulation. J) Statistical analysis of poking currents (n = 6). K) The concentration of calcium ions in stem cells of different groups as detected by flow cytometry (siRNA1: targeting the TRPM4 gene, siRNA2: targeting the PIEZO1 gene). L) Quantitative analysis of flow cytometric results (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).

Article Snippet: TRPC1 inhibitor (0.3 nM, Pico145, CAS No. 1628287-16-0), TRPM7 inhibitor (1.0 μM, VPC4, CAS No. 945604-76-2), TRPV2 inhibitor (5.0 μM, compound IV2-1, CAS No. 2242724-49-6), TRPM4 inhibitor (1.5 μM, CBA, CAS No. 351424-20-9), PIEZO1 inhibitor (2.5 μM, GsMTx4, CAS No. 1209500-46-8), integrin αvβ5 inhibitor (8.0 nM, Compound 12, CAS No.: 2615912-33-7), integrin αvβ1 inhibitor (0.3 nM, Compound C8, CAS No. 1689540-62-2), integrin α5β1 inhibitor (10 μM, ATN-161, 904763-27-5), and CDK5 inhibitor (5 nM, CDK5-IN-1, 2,639,540-19-3) were purchased from MCE Biotechnology Co., LTD. After the MSCs were treated, the cRGD solution was added at a concentration of 1:200 and incubated in the dark for 15 min, and the results were observed by fluorescence microscopy.

Techniques: Activation Assay, Fluorescence, Microscopy, Single Cell, Patch Clamp, Generated, Concentration Assay, Flow Cytometry, Comparison

A) Representative trace from whole-cell patch clamp electrophysiology of a primary rat OPC in culture. The top of the graph is the stimulation trace showing the 300 ms stimulation trace with 0.4 µm step-wise increases in membrane displacement using a glass stimulation rod. The bottom portion of the graph shows the current (pA) trace at each stimulation. 13 of the 14 tested cells showed detectable MA currents. Scale bar: 10 μm B) Maximal current (Imax) for each cell in response to indentation stimulation N = 2 animals, 14 cells, 2 independent cell isolations. The red data point is associated with the representative trace in . C) Inactivation time constant for each cell responsive to indentation stimulation N = 2 animals, 13 cells, 2 independent cell isolations. The red data point is associated with the representative trace in . D) qRT-PCR of Piezo1 , Tmem63a , Mag , and Cspg4 in cultured rat OLs during differentiation (0-192 h). Values are normalized to the transcript’s highest expression value during differentiation N = 3 animals, 3 independent cell isolations. E) Western blot analysis of PIEZO1-tdTOMATO, PDGFRA, MBP, and ACTB expression in Piezo1 tdT/tdT and Piezo1 Wt/Wt control murine OPCs and OLs at 24-72h differentiation. F) Quantification of tdTOMATO intensity relative to ACTB in western blots. N = 3 animals per genotype, 3 independent cell isolations. Red dots indicate the data points corresponding to the example image in E.

Journal: bioRxiv

Article Title: PIEZOs regulate oligodendrocyte sheath formation, expansion, and myelination potential

doi: 10.64898/2026.04.23.720488

Figure Lengend Snippet: A) Representative trace from whole-cell patch clamp electrophysiology of a primary rat OPC in culture. The top of the graph is the stimulation trace showing the 300 ms stimulation trace with 0.4 µm step-wise increases in membrane displacement using a glass stimulation rod. The bottom portion of the graph shows the current (pA) trace at each stimulation. 13 of the 14 tested cells showed detectable MA currents. Scale bar: 10 μm B) Maximal current (Imax) for each cell in response to indentation stimulation N = 2 animals, 14 cells, 2 independent cell isolations. The red data point is associated with the representative trace in . C) Inactivation time constant for each cell responsive to indentation stimulation N = 2 animals, 13 cells, 2 independent cell isolations. The red data point is associated with the representative trace in . D) qRT-PCR of Piezo1 , Tmem63a , Mag , and Cspg4 in cultured rat OLs during differentiation (0-192 h). Values are normalized to the transcript’s highest expression value during differentiation N = 3 animals, 3 independent cell isolations. E) Western blot analysis of PIEZO1-tdTOMATO, PDGFRA, MBP, and ACTB expression in Piezo1 tdT/tdT and Piezo1 Wt/Wt control murine OPCs and OLs at 24-72h differentiation. F) Quantification of tdTOMATO intensity relative to ACTB in western blots. N = 3 animals per genotype, 3 independent cell isolations. Red dots indicate the data points corresponding to the example image in E.

Article Snippet: Piezo1 (ThermoFisher, Rn01432593_m1) Piezo2 (ThermoFisher, Rn01491821_m1) Tmem63a (ThermoFisher, Rn01415187_m1) Mag (ThermoFisher, Rn01457782_m1) Cspg4 (ThermoFisher, Rn00578849_m1) GAPDH (ThermoFisher, Rn0177563_g1)

Techniques: Patch Clamp, Membrane, Quantitative RT-PCR, Cell Culture, Expressing, Western Blot, Control

A. Raw images from Fura2 calcium imaging after the application of DMSO, 30 µM Yoda1, and 200 mM KCl. Scale bar: 25 μm B. Representative calcium imaging trace from control ( Piezo1 Wt/Wt ;Olig2 Cre/Wt ) OPCs loaded with Fura2 and sequentially exposed to DMSO, 30 µM Yoda1, and 200 mM KCl. Trace averaged from 72 cells imaged from 1 coverslip. C. Representative calcium imaging trace from Piezo1 CKO cells ( Piezo1 Fl/Fl ;Olig2 Cre/Wt ) loaded with Fura2 and exposed to DMSO, 30 µM Yoda1, and 200 mM KCl. Trace averaged from 121 CKO cells from a single coverslip. D. Quantification of area under the 340/380 curve for each animal during application of Yoda1. The imaging data from 3 coverslips per animal were pooled for analysis for 3 animals per genotype. Data shown as mean ± SEM. p = 0.024 by student’s t-test, N = 3 animals per genotype, 971 control cells and 1043 cells from Piezo1 CKO cells, 3 independent isolations. E. Representative images from EdU proliferation assay of control or Piezo1 CKO cells. OLIG2 is a marker of OLCs, and EdU labels cells that divided over the 6.5-hour time course. F. Experimental design for the detection of proliferating cells in vitro . G. Quantification of the proportion of OLIG2+ cells labelled for EdU. N = 3 independent OPC isolations per genotype, with 818 cells assessed for controls and 947 cells for Piezo1 CKOs . p = 0.643 by paired t-test. H. Representative images from control or Piezo1 CKO cells fixed 48 hours into differentiation and stained for OLIG2, PDGFRA, and MBP. I. Quantification of the percentage of OLIG2+ cells that are PDGFRA+ OPCs. Control experimental totals: 859 cells, 3 animals, 3 independent OPC isolations CKO experimental totals: 824 cells, 3 animals, 3 independent OPC isolations. p = 0.207 by paired, 2-tailed t-test. J. Quantification of the percentage of OLIG2+ cells that are actively differentiating (PDGFRA-, MBP-). Control experimental totals: 859 cells, 3 animals, 3 independent OPC isolations CKO experimental totals: 824 cells, 3 animals, 3 independent OPC isolations. p = 0.101 by paired, 2-tailed t-test. K. Quantification of the percentage of OLIG2+ cells that are MBP+ OLs. Control experimental totals: 859 cells, 3 animals, 3 independent OPC isolations CKO experimental totals: 824 cells, 3 animals, 3 independent OPC isolations. p = 0.097 by paired, 2-tailed t-test. L. Representative images of a control and Piezo1 CKO cell plated onto nanofibers and allowed to differentiate for 7 days before being fixed and stained with DAPI and MBP. M. The number of sheaths per OL for Control and Piezo1 CKO cells. p = 0.767 by paired, 2-tailed, t-test. N. Average sheath length per OL for Control and Piezo1 CKO cells. p = 0.019 by paired, 2-tailed t-test. O. Average myelin output (summed sheath lengths per OL) for control ( Piezo1 wt/wt ;Olig2 Cre/wt ) and Piezo1 CKO cells. p = 0.032 by paired, 2-tailed t-test. For M-O, Control experimental totals: 50 cells, 3 animals, 3 independent cell isolations. CKO experimental totals: 48 cells, 3 animals, 3 independent cell isolations. For E, H, and L – Scale bar: 50 μm

Journal: bioRxiv

Article Title: PIEZOs regulate oligodendrocyte sheath formation, expansion, and myelination potential

doi: 10.64898/2026.04.23.720488

Figure Lengend Snippet: A. Raw images from Fura2 calcium imaging after the application of DMSO, 30 µM Yoda1, and 200 mM KCl. Scale bar: 25 μm B. Representative calcium imaging trace from control ( Piezo1 Wt/Wt ;Olig2 Cre/Wt ) OPCs loaded with Fura2 and sequentially exposed to DMSO, 30 µM Yoda1, and 200 mM KCl. Trace averaged from 72 cells imaged from 1 coverslip. C. Representative calcium imaging trace from Piezo1 CKO cells ( Piezo1 Fl/Fl ;Olig2 Cre/Wt ) loaded with Fura2 and exposed to DMSO, 30 µM Yoda1, and 200 mM KCl. Trace averaged from 121 CKO cells from a single coverslip. D. Quantification of area under the 340/380 curve for each animal during application of Yoda1. The imaging data from 3 coverslips per animal were pooled for analysis for 3 animals per genotype. Data shown as mean ± SEM. p = 0.024 by student’s t-test, N = 3 animals per genotype, 971 control cells and 1043 cells from Piezo1 CKO cells, 3 independent isolations. E. Representative images from EdU proliferation assay of control or Piezo1 CKO cells. OLIG2 is a marker of OLCs, and EdU labels cells that divided over the 6.5-hour time course. F. Experimental design for the detection of proliferating cells in vitro . G. Quantification of the proportion of OLIG2+ cells labelled for EdU. N = 3 independent OPC isolations per genotype, with 818 cells assessed for controls and 947 cells for Piezo1 CKOs . p = 0.643 by paired t-test. H. Representative images from control or Piezo1 CKO cells fixed 48 hours into differentiation and stained for OLIG2, PDGFRA, and MBP. I. Quantification of the percentage of OLIG2+ cells that are PDGFRA+ OPCs. Control experimental totals: 859 cells, 3 animals, 3 independent OPC isolations CKO experimental totals: 824 cells, 3 animals, 3 independent OPC isolations. p = 0.207 by paired, 2-tailed t-test. J. Quantification of the percentage of OLIG2+ cells that are actively differentiating (PDGFRA-, MBP-). Control experimental totals: 859 cells, 3 animals, 3 independent OPC isolations CKO experimental totals: 824 cells, 3 animals, 3 independent OPC isolations. p = 0.101 by paired, 2-tailed t-test. K. Quantification of the percentage of OLIG2+ cells that are MBP+ OLs. Control experimental totals: 859 cells, 3 animals, 3 independent OPC isolations CKO experimental totals: 824 cells, 3 animals, 3 independent OPC isolations. p = 0.097 by paired, 2-tailed t-test. L. Representative images of a control and Piezo1 CKO cell plated onto nanofibers and allowed to differentiate for 7 days before being fixed and stained with DAPI and MBP. M. The number of sheaths per OL for Control and Piezo1 CKO cells. p = 0.767 by paired, 2-tailed, t-test. N. Average sheath length per OL for Control and Piezo1 CKO cells. p = 0.019 by paired, 2-tailed t-test. O. Average myelin output (summed sheath lengths per OL) for control ( Piezo1 wt/wt ;Olig2 Cre/wt ) and Piezo1 CKO cells. p = 0.032 by paired, 2-tailed t-test. For M-O, Control experimental totals: 50 cells, 3 animals, 3 independent cell isolations. CKO experimental totals: 48 cells, 3 animals, 3 independent cell isolations. For E, H, and L – Scale bar: 50 μm

Techniques: Imaging, Control, Proliferation Assay, Marker, In Vitro, Staining

A. Schematic displaying the features of the knockdown Cas13d targeting system. Tol2 allows integration into the genome, eGFP-CAAX enables morphological characterization of cells, and 4 sites for crRNA guides mediate gene-specific knockdown. B. Experimental timeline starting with embryo harvest on the first day of the experiment, followed by longitudinal imaging of zebrafish embryos at 3, 5, and 7 dpf. C. Representative images of non-targeted control and piezo1-KD cell tracked through time from OL age 1-5, with day 1 being the first day of sheath formation. D. Number of sheaths per OL for non-targeted control and piezo1-KD cells across the experimental timeline. Control: OL1/3: N = 13 cells; 9 fish. OL5: 7 cells; 5 fish. piezo1-KD : OL1/3: N = 12 cells; 7 fish OL5: 9 cells; 6 fish. LMEM (Linear Mixed Effects Model) p cond : 0.002 p cell age:cond :0.009 t-test p OL1 : 0.024, p OL3 : 0.059, p OL5 : 0.039 E. Average sheath length per OL for non-targeted control and piezo1-KD cells. Control: OL1/3: N = 13 cells; 9 fish. OL5: 7 cells; 5 fish. piezo1-KD : OL1/3: N = 12 cells; 7 fish OL5: 9 cells; 6 fish. LMEM p cond : 0.121 p cell age:cond : 0.248 t-test p OL1 : 0.448, p OL3 : 0.194, p OL5 : 0.665 F. Total myelin output per OL for non-targeted control and piezo1-KD cells. Control: OL1/3: N = 13 cells; 9 fish. OL5: 7 cells; 5 fish. piezo1-KD : OL1/3: N = 12 cells; 7 fish OL5: 9 cells; 6 fish. LMEM p cond : 0.123 p cell age:cond :0.234 t-test p OL1 : 0.545, p OL3 : 0.135, p OL5 : 0.052 G. Representative images of non-targeted control and piezo2-KD cells tracked from OL age 1-5, with day 1 being the first day of sheath formation. H. Number of sheaths per OL for non-targeted control and piezo2-KD cells. Control : OL1/3: N = 13 cells; 9 fish. OL5: 12 cells; 8 fish. piezo2-KD : OL1/3: N = 14 cells; 10 fish OL5: 9 cells; 8 fish. LMEM p cond : 0.046 p cell age:cond : 0.371 t-test p OL1 : 0.981, p OL3 : 0.456, p OL5 : <0.001 I. Average sheath length per OL for non-targeted control and piezo2-KD cells. Control : OL1/3: N = 13 cells; 9 fish. OL5: 12 cells; 8 fish. piezo2-KD : OL1/3: N = 14 cells; 10 fish OL5: 9 cells; 8 fish. LMEM p cond : 0.207 p cell age:cond : 0.377 t-test p OL1 : 0.364, p OL3 : 0.850, p OL5 : 0.669 J. Total myelin output per OL for non-targeted controls and piezo2-KD cells. Control : OL1/3: N = 13 cells; 9 fish. OL5: 12 cells; 8 fish. piezo2-KD : OL1/3: N = 14 cells; 10 fish OL5: 9 cells; 8 fish. LMEM p cond : 0.634 p cell age:cond : 0.033 t-test p OL1 : 0.864, p OL3 : 0.115, p OL5 : 0.003 For C/G – Scale bar: 25 μm. For E-F and H-J graphs are mean ± SEM and * represent statistically significant timepoints.

Journal: bioRxiv

Article Title: PIEZOs regulate oligodendrocyte sheath formation, expansion, and myelination potential

doi: 10.64898/2026.04.23.720488

Figure Lengend Snippet: A. Schematic displaying the features of the knockdown Cas13d targeting system. Tol2 allows integration into the genome, eGFP-CAAX enables morphological characterization of cells, and 4 sites for crRNA guides mediate gene-specific knockdown. B. Experimental timeline starting with embryo harvest on the first day of the experiment, followed by longitudinal imaging of zebrafish embryos at 3, 5, and 7 dpf. C. Representative images of non-targeted control and piezo1-KD cell tracked through time from OL age 1-5, with day 1 being the first day of sheath formation. D. Number of sheaths per OL for non-targeted control and piezo1-KD cells across the experimental timeline. Control: OL1/3: N = 13 cells; 9 fish. OL5: 7 cells; 5 fish. piezo1-KD : OL1/3: N = 12 cells; 7 fish OL5: 9 cells; 6 fish. LMEM (Linear Mixed Effects Model) p cond : 0.002 p cell age:cond :0.009 t-test p OL1 : 0.024, p OL3 : 0.059, p OL5 : 0.039 E. Average sheath length per OL for non-targeted control and piezo1-KD cells. Control: OL1/3: N = 13 cells; 9 fish. OL5: 7 cells; 5 fish. piezo1-KD : OL1/3: N = 12 cells; 7 fish OL5: 9 cells; 6 fish. LMEM p cond : 0.121 p cell age:cond : 0.248 t-test p OL1 : 0.448, p OL3 : 0.194, p OL5 : 0.665 F. Total myelin output per OL for non-targeted control and piezo1-KD cells. Control: OL1/3: N = 13 cells; 9 fish. OL5: 7 cells; 5 fish. piezo1-KD : OL1/3: N = 12 cells; 7 fish OL5: 9 cells; 6 fish. LMEM p cond : 0.123 p cell age:cond :0.234 t-test p OL1 : 0.545, p OL3 : 0.135, p OL5 : 0.052 G. Representative images of non-targeted control and piezo2-KD cells tracked from OL age 1-5, with day 1 being the first day of sheath formation. H. Number of sheaths per OL for non-targeted control and piezo2-KD cells. Control : OL1/3: N = 13 cells; 9 fish. OL5: 12 cells; 8 fish. piezo2-KD : OL1/3: N = 14 cells; 10 fish OL5: 9 cells; 8 fish. LMEM p cond : 0.046 p cell age:cond : 0.371 t-test p OL1 : 0.981, p OL3 : 0.456, p OL5 : <0.001 I. Average sheath length per OL for non-targeted control and piezo2-KD cells. Control : OL1/3: N = 13 cells; 9 fish. OL5: 12 cells; 8 fish. piezo2-KD : OL1/3: N = 14 cells; 10 fish OL5: 9 cells; 8 fish. LMEM p cond : 0.207 p cell age:cond : 0.377 t-test p OL1 : 0.364, p OL3 : 0.850, p OL5 : 0.669 J. Total myelin output per OL for non-targeted controls and piezo2-KD cells. Control : OL1/3: N = 13 cells; 9 fish. OL5: 12 cells; 8 fish. piezo2-KD : OL1/3: N = 14 cells; 10 fish OL5: 9 cells; 8 fish. LMEM p cond : 0.634 p cell age:cond : 0.033 t-test p OL1 : 0.864, p OL3 : 0.115, p OL5 : 0.003 For C/G – Scale bar: 25 μm. For E-F and H-J graphs are mean ± SEM and * represent statistically significant timepoints.

Techniques: Knockdown, Imaging, Control

A. Example tracing of OLs used to quantify the number of sheaths per OL for non-targeted control and double knockdown (dKD) cells tracked from OL1-5 (3 to 7 dpf). Arrowhead indicates the formation of a new sheath generated outside the normal developmental window. B. Change in the number of sheaths per OL between 5 to 7 dpf for each condition. The targeting of piezo1 , piezo2 , and dKD were performed as independent experiments; therefore, corresponding control groups are shown for each experiment. Control P1 : N = 7 cells, 7 animals; piezo1-KD : N = 11 cells; 8 fish; p = 0.536 Control P2 : N = 13 cells, 10 animals; piezo2-KD : N = 9 cells, 8 fish; p = 0.781 Control dKD :11 cells, 11 fish; dKD: 10 cells, 8 fish; p = 0.014

Journal: bioRxiv

Article Title: PIEZOs regulate oligodendrocyte sheath formation, expansion, and myelination potential

doi: 10.64898/2026.04.23.720488

Figure Lengend Snippet: A. Example tracing of OLs used to quantify the number of sheaths per OL for non-targeted control and double knockdown (dKD) cells tracked from OL1-5 (3 to 7 dpf). Arrowhead indicates the formation of a new sheath generated outside the normal developmental window. B. Change in the number of sheaths per OL between 5 to 7 dpf for each condition. The targeting of piezo1 , piezo2 , and dKD were performed as independent experiments; therefore, corresponding control groups are shown for each experiment. Control P1 : N = 7 cells, 7 animals; piezo1-KD : N = 11 cells; 8 fish; p = 0.536 Control P2 : N = 13 cells, 10 animals; piezo2-KD : N = 9 cells, 8 fish; p = 0.781 Control dKD :11 cells, 11 fish; dKD: 10 cells, 8 fish; p = 0.014

Techniques: Control, Knockdown, Generated

Journal: Biomaterials

Article Title: A comparative study assessing neural recording quality and inflammatory tissue response between stiff and flexible microelectrode arrays

doi: 10.1016/j.biomaterials.2025.123929

Figure Lengend Snippet: Heightened piezo1 expression around the stiff implant. A) Representative images showing Piezo1 expression around the flexible (left) and stiff (right) implant site (*). B) Quantification of normalized intensity as a function of distance from the implant (μm). C) Normalized intensity averaged across the first 50 μm from the implant site. 19 images for the flexible and 22 images for the stiff group were analyzed. D) Representative images showing Piezo1 (cyan), Iba-1 (green), GFAP (red), and merged expression around a stiff implant, with the zoomed inset with DAPI (blue) overlay. E) Correlation of total piezo1 expression with total Iba-1 expression per image for flexible (left, blue) and stiff (right, red) groups. The slope is significant for the stiff group, indicating a strong correlation between Piezo1 expression and Iba-1 intensity. F) Correlation of total piezo1 expression with total GFAP expression per image for flexible (left, blue) and stiff (right, red). 4 animals were implanted in each group. C) Two-way ANOVA with Sidak’s multiple comparison test was used, E-F) Data was normalized per image and the mean set to 0. A simple linear regression was performed, dashed lines indicate 95 % confidence intervals, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. The scale bar is 100 μm in A) and D) and 50 μm in the zoomed inset in D). C) Data are mean ± SEM. E-F) Data is sum.

Article Snippet: Antibodies to visualize astrocytes (GFAP, 1:500, Z033401 Dako), macrophage/microglia IBA-1(microglia, 1:500, NC9288364 Fisher), Piezo1 mechanosensitive ion channels (Piezo1,1:100, NBP275617 Novus Biologicals), neuronal nuclei (NeuN, 1:750, CAD69 Abcam), and neural filament (NF200, 1:250 MAB5256 Millipore) were used.

Techniques: Expressing, Comparison

Journal: Biomaterials

Article Title: A comparative study assessing neural recording quality and inflammatory tissue response between stiff and flexible microelectrode arrays

doi: 10.1016/j.biomaterials.2025.123929

Figure Lengend Snippet: Cell-specific intensity analysis. A) Microglia mask of a sample image using Iba-1. Black portions represent the extracted cell structures. This mask was used to identify the sample microglia shown in B). B) Piezo1 (blue) stained image showing activated microglia expressing Piezo1. C) Average within-microglia Piezo1 intensity with distance from the probe center. D) Average within-microglia Iba-1 intensity with distance from the probe center. E) Astrocyte mask of the above image using GFAP. Black portions represent the extracted cell structures. This mask was used to identify the sample astrocyte processes shown in F). F) Piezo1 (blue) stained image showing astrocyte processes expressing Piezo1. G) Average within-astrocyte Piezo1 intensity with distance from the probe center. H) Average within-astrocyte GFAP intensity with distance from the probe center. For all microglia and astrocyte analysis, the increase in intensity with proximity to the injury is highly significant for both probe substrates, and the cells surrounding stiff probes have higher intensity expression than the flexible. I) Piezo1 (red) stained image overlayed with NeuN ROIs produced by Cellpose. J) Piezo1 (red) stained image merged with NeuN (green). K) Average within-neuron Piezo1 intensity with distance from the probe center. Sample DAPI (white) stained image for a L) stiff and M) flexible probe implant. N) Average DAPI circularity index for all images analyzed above. Bar plot is the same data, but binned into two large bins rather than 10 evenly spaced bins. For A) and E), DAPI is overlayed on the mask. DAPI that passes through masked cell structures is seen as red, while DAPI covered by the mask is seen as yellow. Scale bar is 100 μm and 50 μm in zoomed insets in M&N. * Indicates implantation site. For comparisons between stiff and flexible, a two-way ANOVA with Sidak’s multiple comparison test was used. For correlation of intensity with distance from the probe, a Spearman’s r correlation test was used due to the non-normality of many of the datasets, **p < 0.01, ***p < 0.001, ****p < 0.0001. All data is mean ± SEM.

Techniques: Staining, Expressing, Produced, Comparison

Piezo1 expression is upregulated in the lung epithelium during ventilator-induced lung injury. (A) Representative immunofluorescence images of lung sections from sham-operated control mice and mice subjected to 6 h of mechanical ventilation. Piezo1 protein (red) is constitutively expressed and colocalizes (yellow in merged panels) with the epithelial cell marker CK8 (green). Nuclei were counterstained with DAPI (blue). Scale bar, 100 µm. (B) Representative images of immunohistochemical staining for Piezo1 expression in lung epithelium from sham-operated mice and mice subjected to 6 h of mechanical ventilation. Scale bar, 20 µm (original magnification, ×400). (C) Representative haematoxylin and eosin-stained lung sections from sham-operated and HTV-ventilated mice. Scale bar, 50 µm. (D) Quantitative lung injury score was determined based on histological evaluation of alveolar congestion, haemorrhage, leukocyte infiltration and alveolar wall thickness, each graded from 0 (normal) to 3 (severe). Data are presented as the median (interquartile range); n=6 mice/group and Pvalues were determined by Mann-Whitney U test. Lung injury was further evaluated on the basis of (E) protein concentration, (F) cell number in BALF and (G) wet/dry weight ratio. (H) MPO activity in lung tissue. Proinflammatory cytokines in BALF were evaluated using the following ELISA kits: (I) IL-6, (J) TNF-α and (K) IL-β. (E-K) Data are presented as the mean ± SD, n=6 mice/group. Pvalues were determined by Student's t-test. *P<0.05, **P<0.01, ***P<0.001. BALF, bronchoalveolar lavage fluid; CK8, cytokeratin 8; HTV, high tidal volume; MPO, myeloperoxidase.

Journal: Molecular Medicine Reports

Article Title: Mechanotransducer Piezo1 drives ventilator-induced lung injury in lung epithelial cells via the calcineurin/NFATc3 pathway

doi: 10.3892/mmr.2026.13893

Figure Lengend Snippet: Piezo1 expression is upregulated in the lung epithelium during ventilator-induced lung injury. (A) Representative immunofluorescence images of lung sections from sham-operated control mice and mice subjected to 6 h of mechanical ventilation. Piezo1 protein (red) is constitutively expressed and colocalizes (yellow in merged panels) with the epithelial cell marker CK8 (green). Nuclei were counterstained with DAPI (blue). Scale bar, 100 µm. (B) Representative images of immunohistochemical staining for Piezo1 expression in lung epithelium from sham-operated mice and mice subjected to 6 h of mechanical ventilation. Scale bar, 20 µm (original magnification, ×400). (C) Representative haematoxylin and eosin-stained lung sections from sham-operated and HTV-ventilated mice. Scale bar, 50 µm. (D) Quantitative lung injury score was determined based on histological evaluation of alveolar congestion, haemorrhage, leukocyte infiltration and alveolar wall thickness, each graded from 0 (normal) to 3 (severe). Data are presented as the median (interquartile range); n=6 mice/group and Pvalues were determined by Mann-Whitney U test. Lung injury was further evaluated on the basis of (E) protein concentration, (F) cell number in BALF and (G) wet/dry weight ratio. (H) MPO activity in lung tissue. Proinflammatory cytokines in BALF were evaluated using the following ELISA kits: (I) IL-6, (J) TNF-α and (K) IL-β. (E-K) Data are presented as the mean ± SD, n=6 mice/group. Pvalues were determined by Student's t-test. *P<0.05, **P<0.01, ***P<0.001. BALF, bronchoalveolar lavage fluid; CK8, cytokeratin 8; HTV, high tidal volume; MPO, myeloperoxidase.

Article Snippet: To establish male mice with tamoxifen-inducible, lung epithelial-specific deletion of Piezo1 ( Piezo1 CKO ), Piezo1 F/F mice ( Piezo1 tm2.1Apat/ J; Jackson Laboratory) were bred with Sftpc-Cre ERT mice (Jackson Laboratory) in our laboratory.

Techniques: Expressing, Immunofluorescence, Control, Marker, Immunohistochemical staining, Staining, MANN-WHITNEY, Protein Concentration, Activity Assay, Enzyme-linked Immunosorbent Assay

CS induces Piezo1 expression and epithelial injury markers in lung epithelial cells in vitro . (A) Representative western blot analysis of Piezo1 protein expression in lung epithelial cells under control (unstretched) conditions, and after 3 or 6 h of CS. (B) Semi-quantification of Piezo1 protein expression normalized to that of GAPDH. (C) Piezo1 mRNA expression in sham and stretched cells (after 3 and 6 h). (D) Representative fluorescence images of F-actin (phalloidin, green) and nuclei (DAPI, blue) in control and stretched cells. Scale bar, 20 µm. Proinflammatory cytokine levels of (E) IL-6, (F) TNF-α and (G) IL-1β protein levels in sham versus stretched cells. Data are presented as the mean ± SD, n=6 each. Pvalues were determined by oneway ANOVA. *P<0.05, **P<0.01, ***P<0.001. CS, cyclic stretch.

Journal: Molecular Medicine Reports

Article Title: Mechanotransducer Piezo1 drives ventilator-induced lung injury in lung epithelial cells via the calcineurin/NFATc3 pathway

doi: 10.3892/mmr.2026.13893

Figure Lengend Snippet: CS induces Piezo1 expression and epithelial injury markers in lung epithelial cells in vitro . (A) Representative western blot analysis of Piezo1 protein expression in lung epithelial cells under control (unstretched) conditions, and after 3 or 6 h of CS. (B) Semi-quantification of Piezo1 protein expression normalized to that of GAPDH. (C) Piezo1 mRNA expression in sham and stretched cells (after 3 and 6 h). (D) Representative fluorescence images of F-actin (phalloidin, green) and nuclei (DAPI, blue) in control and stretched cells. Scale bar, 20 µm. Proinflammatory cytokine levels of (E) IL-6, (F) TNF-α and (G) IL-1β protein levels in sham versus stretched cells. Data are presented as the mean ± SD, n=6 each. Pvalues were determined by oneway ANOVA. *P<0.05, **P<0.01, ***P<0.001. CS, cyclic stretch.

Techniques: Expressing, In Vitro, Western Blot, Control, Fluorescence

Lung epithelial-specific Piezo1 deletion attenuates ventilator-induced lung injury. (A) Validation of Piezo1 knockout efficiency in the lung epithelium. Representative immunofluorescence images of lung sections from control and Piezo1 CKO mice showing the expression of the Piezo1 protein (red), the epithelial cell marker CK8 (green), and nuclei (DAPI, blue). Scale bar, 50 µm. (B) Representative haematoxylin and eosin-stained lung sections from control and Piezo1 CKO mice after 6 h of mechanical ventilation. Scale bar, 50 µm. (C) Quantitative histopathological lung injury score. Data are presented as the median (IQR); n=6/group. Data were analysed using the Kruskal-Wallis test followed by Dunn's post hoc test. (D) Total protein concentration in BALF. (E) Total cell counts in the BALF. (F) Lung wet/dry weight ratio. (G) Lung MPO activity. BALF concentrations of the inflammatory cytokines (H) IL-6, (I) TNF-α and (J) IL-1β. (D-J) Data are presented as the mean ± SD, n=6 mice/group. Pvalues were determined by oneway ANOVA. *P<0.05, **P<0.01, ***P<0.001. BALF, bronchoalveolar lavage fluid; CK8, cytokeratin 8; CKO, conditional; HTV, high tidal volume; MPO, myeloperoxidase.

Journal: Molecular Medicine Reports

Article Title: Mechanotransducer Piezo1 drives ventilator-induced lung injury in lung epithelial cells via the calcineurin/NFATc3 pathway

doi: 10.3892/mmr.2026.13893

Figure Lengend Snippet: Lung epithelial-specific Piezo1 deletion attenuates ventilator-induced lung injury. (A) Validation of Piezo1 knockout efficiency in the lung epithelium. Representative immunofluorescence images of lung sections from control and Piezo1 CKO mice showing the expression of the Piezo1 protein (red), the epithelial cell marker CK8 (green), and nuclei (DAPI, blue). Scale bar, 50 µm. (B) Representative haematoxylin and eosin-stained lung sections from control and Piezo1 CKO mice after 6 h of mechanical ventilation. Scale bar, 50 µm. (C) Quantitative histopathological lung injury score. Data are presented as the median (IQR); n=6/group. Data were analysed using the Kruskal-Wallis test followed by Dunn's post hoc test. (D) Total protein concentration in BALF. (E) Total cell counts in the BALF. (F) Lung wet/dry weight ratio. (G) Lung MPO activity. BALF concentrations of the inflammatory cytokines (H) IL-6, (I) TNF-α and (J) IL-1β. (D-J) Data are presented as the mean ± SD, n=6 mice/group. Pvalues were determined by oneway ANOVA. *P<0.05, **P<0.01, ***P<0.001. BALF, bronchoalveolar lavage fluid; CK8, cytokeratin 8; CKO, conditional; HTV, high tidal volume; MPO, myeloperoxidase.

Techniques: Biomarker Discovery, Knock-Out, Immunofluorescence, Control, Expressing, Marker, Staining, Protein Concentration, Activity Assay

Genetic inhibition of Piezo1 attenuates mechanical stretch-induced cytoskeletal disruption and the inflammatory response in lung epithelial cells in vitro . (A) Representative western blotting and (B) semi-quantitative analysis demonstrating efficient knockdown of Piezo1 protein expression in MLE-12 cells transfected with shPiezo1 compared with that in cells transfected with Scr-shRNA. (C) Piezo1 mRNA expression levels measured by quantitative PCR in cells transfected with shPiezo1 compared with those in cells transfected with Scr-shRNA. Proinflammatory cytokine levels in cell culture supernatants from Scr-shRNA and shPiezo1 cells under control conditions or after 6 h of CS: (D) IL-6, (E) TNF-α and (F) IL-1β protein levels. (G) Representative fluorescence images showing F-actin morphology (phalloidin, green) and nuclei (DAPI, blue) in Scr-shPiezo1 and shPiezo1 cells under control (unstretched) conditions or after 6 h of CS. Scale bar, 20 µm. Data are presented as the mean ± SD, n=6 each. Pvalues were determined by oneway ANOVA. **P<0.01, ***P<0.001. CS, cyclic stretch; Scr, scrambled; sh, short hairpin.

Journal: Molecular Medicine Reports

Article Title: Mechanotransducer Piezo1 drives ventilator-induced lung injury in lung epithelial cells via the calcineurin/NFATc3 pathway

doi: 10.3892/mmr.2026.13893

Figure Lengend Snippet: Genetic inhibition of Piezo1 attenuates mechanical stretch-induced cytoskeletal disruption and the inflammatory response in lung epithelial cells in vitro . (A) Representative western blotting and (B) semi-quantitative analysis demonstrating efficient knockdown of Piezo1 protein expression in MLE-12 cells transfected with shPiezo1 compared with that in cells transfected with Scr-shRNA. (C) Piezo1 mRNA expression levels measured by quantitative PCR in cells transfected with shPiezo1 compared with those in cells transfected with Scr-shRNA. Proinflammatory cytokine levels in cell culture supernatants from Scr-shRNA and shPiezo1 cells under control conditions or after 6 h of CS: (D) IL-6, (E) TNF-α and (F) IL-1β protein levels. (G) Representative fluorescence images showing F-actin morphology (phalloidin, green) and nuclei (DAPI, blue) in Scr-shPiezo1 and shPiezo1 cells under control (unstretched) conditions or after 6 h of CS. Scale bar, 20 µm. Data are presented as the mean ± SD, n=6 each. Pvalues were determined by oneway ANOVA. **P<0.01, ***P<0.001. CS, cyclic stretch; Scr, scrambled; sh, short hairpin.

Techniques: Inhibition, Disruption, In Vitro, Western Blot, Knockdown, Expressing, Transfection, shRNA, Real-time Polymerase Chain Reaction, Cell Culture, Control, Fluorescence

Piezo1 mediates CS-induced Ca 2+ influx in epithelial cells. (A) Quantitative analysis of cytosolic Ca 2+ levels (ΔF/F 0 ) measured using a microplate reader in Fluo-3 AM-loaded MLE-12 cells treated with the Piezo1 agonist Yoda1 (10 µM) for 0, 15 and 30 min. (B) Representative fluorescence microscopy images of Fluo-3 AM-loaded MLE-12 cells at 30 min following the onset of CS Scale bar, 50 µm. (C and D) Both genetic knockdown of Piezo1 (with shPiezo1) and pharmacological inhibition with GsMTx4 (5 µM) attenuated the Ca 2+ elevation induced by 30-min CS. (E) Genetic knockdown of Piezo1 (with shPiezo1) attenuated mechanical stretch-induced calcineurin activation after 6 h. (F-H) Pharmacological inhibition of calcineurin with CsA (10 µM) or FK506 (10 µM) significantly reduced mechanical stretch -induced cytokine production after 6 h. (I) Disruption of the F-actin network induced by 6-h mechanical stretching was reversed by pretreatment with CsA or FK506. Scale bar, 20 µm. Data are presented as the mean ± SD, n=6 each. Pvalues were determined by oneway ANOVA. *P<0.05, ***P<0.001. CS, cyclic stretch; CsA, cyclosporine A; Scr, scrambled; sh, short hairpin.

Journal: Molecular Medicine Reports

Article Title: Mechanotransducer Piezo1 drives ventilator-induced lung injury in lung epithelial cells via the calcineurin/NFATc3 pathway

doi: 10.3892/mmr.2026.13893

Figure Lengend Snippet: Piezo1 mediates CS-induced Ca 2+ influx in epithelial cells. (A) Quantitative analysis of cytosolic Ca 2+ levels (ΔF/F 0 ) measured using a microplate reader in Fluo-3 AM-loaded MLE-12 cells treated with the Piezo1 agonist Yoda1 (10 µM) for 0, 15 and 30 min. (B) Representative fluorescence microscopy images of Fluo-3 AM-loaded MLE-12 cells at 30 min following the onset of CS Scale bar, 50 µm. (C and D) Both genetic knockdown of Piezo1 (with shPiezo1) and pharmacological inhibition with GsMTx4 (5 µM) attenuated the Ca 2+ elevation induced by 30-min CS. (E) Genetic knockdown of Piezo1 (with shPiezo1) attenuated mechanical stretch-induced calcineurin activation after 6 h. (F-H) Pharmacological inhibition of calcineurin with CsA (10 µM) or FK506 (10 µM) significantly reduced mechanical stretch -induced cytokine production after 6 h. (I) Disruption of the F-actin network induced by 6-h mechanical stretching was reversed by pretreatment with CsA or FK506. Scale bar, 20 µm. Data are presented as the mean ± SD, n=6 each. Pvalues were determined by oneway ANOVA. *P<0.05, ***P<0.001. CS, cyclic stretch; CsA, cyclosporine A; Scr, scrambled; sh, short hairpin.

Techniques: Fluorescence, Microscopy, Knockdown, Inhibition, Activation Assay, Disruption

$Piezo1 mediates CS-induced inflammation via calcineurin/NFATc3 signalling. (A) Immunofluorescence screening of the nuclear translocation of NFAT isoforms (NFATc1-4) upon CS. NFATc3 showed the most prominent nuclear translocation. Scale bar, 10 µm. (B) Representative western blot images showing NFATc3 protein levels in the nuclear and cytoplasmic fractions of MLE-12 cells transfected with Scr-shRNA or shPiezo1, with or without CS (6 h). GAPDH and Lamin B1 were used as loading controls for cytoplasmic and nuclear fractions, respectively. (C) Semi-quantitative analysis of cytoplasmic NFATc3 protein expression normalized to GAPDH. (D) Semi-quantitative analysis of nuclear NFATc3 protein expression normalized to Lamin B1. (E) Representative western blot images showing NFATc3 protein levels in the nuclear and cytoplasmic fractions of MLE-12 cells treated with or without calcineurin inhibitors (CsA, 10 µM; FK506, 10 µM) prior to CS (6 h). GAPDH and Lamin B1 were used as loading controls for cytoplasmic and nuclear fractions, respectively. (F) Semi-quantitative analysis of cytoplasmic NFATc3 protein expression normalized to GAPDH. (G) Semi-quantitative analysis of nuclear NFATc3 protein expression normalized to Lamin B1. Data are presented as the mean ± SD, n=6 each. Pvalues were determined by oneway ANOVA. *P<0.05, **P<0.01, ***P<0.001. CS, cyclic stretch; CsA, cyclosporine A; Scr, scrambled; sh, short hairpin.$

Journal: Molecular Medicine Reports

Article Title: Mechanotransducer Piezo1 drives ventilator-induced lung injury in lung epithelial cells via the calcineurin/NFATc3 pathway

doi: 10.3892/mmr.2026.13893

Figure Lengend Snippet: Piezo1 mediates CS-induced inflammation via calcineurin/NFATc3 signalling. (A) Immunofluorescence screening of the nuclear translocation of NFAT isoforms (NFATc1-4) upon CS. NFATc3 showed the most prominent nuclear translocation. Scale bar, 10 µm. (B) Representative western blot images showing NFATc3 protein levels in the nuclear and cytoplasmic fractions of MLE-12 cells transfected with Scr-shRNA or shPiezo1, with or without CS (6 h). GAPDH and Lamin B1 were used as loading controls for cytoplasmic and nuclear fractions, respectively. (C) Semi-quantitative analysis of cytoplasmic NFATc3 protein expression normalized to GAPDH. (D) Semi-quantitative analysis of nuclear NFATc3 protein expression normalized to Lamin B1. (E) Representative western blot images showing NFATc3 protein levels in the nuclear and cytoplasmic fractions of MLE-12 cells treated with or without calcineurin inhibitors (CsA, 10 µM; FK506, 10 µM) prior to CS (6 h). GAPDH and Lamin B1 were used as loading controls for cytoplasmic and nuclear fractions, respectively. (F) Semi-quantitative analysis of cytoplasmic NFATc3 protein expression normalized to GAPDH. (G) Semi-quantitative analysis of nuclear NFATc3 protein expression normalized to Lamin B1. Data are presented as the mean ± SD, n=6 each. Pvalues were determined by oneway ANOVA. *P<0.05, **P<0.01, ***P<0.001. CS, cyclic stretch; CsA, cyclosporine A; Scr, scrambled; sh, short hairpin.

Techniques: Immunofluorescence, Translocation Assay, Western Blot, Transfection, shRNA, Expressing

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