polyclonal rabbit anti piezo1  (Novus Biologicals)

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: The Piezo1 expression profile in ovary tumor, breast tumor, brain tumor, liver tumor, ovary tumor, lung tumor and matched healthy tissues, based on analysis of GENT2 datasets. Data were presented as the mean ± SD. ∗∗∗P < 0.001.

Article Snippet: The following primary antibodies were used: rabbit anti-Piezo1 antibody (1:100 dilution, #DF12083, Affinity Biosciences, China) and rabbit anti-β-actin antibody (1:500 dilution, #GB11001-100, Servicebio, China), anti-p-cofilin and anti-cofilin antibodies (1:100 dilution, #SC365882 and #SC376476, Santa, USA).

Techniques: Expressing

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Stiff substrate promotes A549 and H460 cell migration and down-regulates Piezo1 channel e xpression. (A–D) Transwell assay of the effects of substrate stiffness on cell migration. Representative images of migrated cells stained with crystal violet (10x, A-B) and statistical analysis of data from three independent experiments (C–D). Scale bar: 50 μm. (E–H) Flow cytometry assessing the effects of substrate stiffness on cell surface Piezo1 protein expression. Representative images of flow cytometry (E–F) and statistical analysis of data from three (G–F) independent experiments. All data were normalized to that of 3 kPa group. Data were presented as mean ± SD. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗ P < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following primary antibodies were used: rabbit anti-Piezo1 antibody (1:100 dilution, #DF12083, Affinity Biosciences, China) and rabbit anti-β-actin antibody (1:500 dilution, #GB11001-100, Servicebio, China), anti-p-cofilin and anti-cofilin antibodies (1:100 dilution, #SC365882 and #SC376476, Santa, USA).

Techniques: Migration, Transwell Assay, Staining, Flow Cytometry, Expressing

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Piezo1 channel negatively regulates substrate stiffness-induced A549 cell migration. (A, D) Piezo1 channel blockade with GsMTx4 promotes cell migration on both soft and stiff substrates. (B, E) Piezo1 channel activation with Yoda 1 inhibits cell migration on both soft and stiff substrates. (C, F) Piezo1 channel knockdown with specific siRNA transfection promotes cell migration on both soft and stiff substrates. Representative images of migrated cells stained with crystal violet (10x, A-C) and statistical analysis of data from three independent experiments (D–F). Scale bar: 50 μm. All data were normalized to the 3 kPa group. Data were presented as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following primary antibodies were used: rabbit anti-Piezo1 antibody (1:100 dilution, #DF12083, Affinity Biosciences, China) and rabbit anti-β-actin antibody (1:500 dilution, #GB11001-100, Servicebio, China), anti-p-cofilin and anti-cofilin antibodies (1:100 dilution, #SC365882 and #SC376476, Santa, USA).

Techniques: Migration, Activation Assay, Knockdown, Transfection, Staining

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Piezo1 channel negatively regulates stiff substrate-induced filopodia formation in A 549 cells. (A, C, D) Piezo1 channel blockade with GsMTx4 further promotes filopodia formation in cells on both soft and stiff substrates. (B, E, F) Piezo1 channel activation with Yoda 1 further inhibits filopodia formation in cells on stiff substrates but has no effect in cells on soft substrates. Representative images of filopodia morphology (A and B) and statistical analysis of the filopodia length (C and E) and number (D and F) from indicated number of cells. Red, F‐actin staining with rhodamine-labeled phalloidin; blue, nucleus staining with Hoechst 33342. All data were normalized to that of the 3 kPa group. Scale bar: 20 μm. Data were presented as mean ± SD. ∗ P < 0.05; ∗∗∗ P < 0.001; ns, not significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following primary antibodies were used: rabbit anti-Piezo1 antibody (1:100 dilution, #DF12083, Affinity Biosciences, China) and rabbit anti-β-actin antibody (1:500 dilution, #GB11001-100, Servicebio, China), anti-p-cofilin and anti-cofilin antibodies (1:100 dilution, #SC365882 and #SC376476, Santa, USA).

Techniques: Activation Assay, Staining, Labeling

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Piezo1 channel mediates substrate stiffness-induced change in [Ca 2+ ] i in A 549 cells. (A, B) Cells showed the higher and lower [Ca 2+ ] i in cells on soft and stiff substrates, respectively. (C, D) Piezo1 channel blockade with GsMTx4 reduces [Ca 2+ ] i in cells on both soft and stiff substrates. Representative Ca 2+ images (A, C and E) and statistical analysis of [Ca 2+ ] i in indicated numbers of cells (B and D). Scale bar: 50 μm. All data were normalized to that of 3 kPa group. Data were presented as mean ± SD. ∗∗∗ P < 0.001; ns, not significant.

Article Snippet: The following primary antibodies were used: rabbit anti-Piezo1 antibody (1:100 dilution, #DF12083, Affinity Biosciences, China) and rabbit anti-β-actin antibody (1:500 dilution, #GB11001-100, Servicebio, China), anti-p-cofilin and anti-cofilin antibodies (1:100 dilution, #SC365882 and #SC376476, Santa, USA).

Techniques:

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Piezo1 channel mediates a strong but weak calcium influx induced by soft and stiff substrates, respectively. (A and B) Extrcellular Ca 2+ influx in cells on soft and stiff substrates. (C and D) Piezo1 blockade with GsMTx4 abolished the difference in Ca 2+ influx between cells on soft and stiff substrates. Representative tracces showing change in [Ca 2+ ] i (A and C) and statistical analysis of the maximal change in [Ca 2+ ] i in indicated numbers of cells (B and D). Cells were cultured in medium with or without 2.5 μM GsMTx4 containment for 48 h. Cells loaded with Fluo4-AM were imaged with 5 s interval in Ca 2+ -free buffer for 1 min and further 4 min upon addition of 2 mM CaCl 2 . Scale bar: 50 μm. All data were normalized to that ones prior to addition of CaCl 2 . Data were presented as mean ± SD. ∗∗∗ P < 0.001.

Article Snippet: The following primary antibodies were used: rabbit anti-Piezo1 antibody (1:100 dilution, #DF12083, Affinity Biosciences, China) and rabbit anti-β-actin antibody (1:500 dilution, #GB11001-100, Servicebio, China), anti-p-cofilin and anti-cofilin antibodies (1:100 dilution, #SC365882 and #SC376476, Santa, USA).

Techniques: Cell Culture

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Piezo1 channel regulates stiff substrate-induced phosphorylation of coflilin through reducing the [Ca 2+ ] i in A 549 cells. (A–C) Stiff substrate induces phosphorylation of cofilin (C), without effect on its expression (B). (D–G) Stiff substrate-induced phosphorylation of coflilin is enhanced by Piezo1 channel blockade with GsMTx4 (D and E) but attenuated by Piezo1 channel activation with Yoda-1 (F and G). (H, I) Chelation of intracellular Ca 2+ with BAPTA-AM promotes cofilin phosphorylation in cells on both soft and stiff substrates. Representative images of western blotting (A, D, F and H) and statistical analysis of data from three independent experiments (B, C, E, G and I). All data were normalized to that of the 3 kPa group. Data are presented as mean ± SD. ∗ P < 0.05; ∗ P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: The following primary antibodies were used: rabbit anti-Piezo1 antibody (1:100 dilution, #DF12083, Affinity Biosciences, China) and rabbit anti-β-actin antibody (1:500 dilution, #GB11001-100, Servicebio, China), anti-p-cofilin and anti-cofilin antibodies (1:100 dilution, #SC365882 and #SC376476, Santa, USA).

Techniques: Phospho-proteomics, Expressing, Activation Assay, Western Blot

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Piezo1 channel regulates stiff substrate-induced phosphorylation of coflilin through attenuating the Ca 2+ -dependent CaN/SSH activation in A 549 cells. (A–B) CaN inhibition with CsA promotes cofilin phosphorylation in cells on both soft and stiff substrates. (C–D) CaN activity was decreased on stiff substrate, and further decreased on soft and stiff substrates by Piezo1 channel blockade with GsMTx4 (B) or Chelation of intracellular Ca 2+ with BAPTA-AM (D), respectivley. (E–F) The p-SSH1 was incrased on stiff substrates, and further increased on soft and stiff substrates by CaN inhibition with CsA. Representative images of western blotting (A) and flow cytometry (E), and statistical analysis of data from three independent experiments (B, C, D and F). All data were normalized to that of 3 kPa group. Data are presented as mean ± SD. ∗ P < 0.05; ∗ P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: The following primary antibodies were used: rabbit anti-Piezo1 antibody (1:100 dilution, #DF12083, Affinity Biosciences, China) and rabbit anti-β-actin antibody (1:500 dilution, #GB11001-100, Servicebio, China), anti-p-cofilin and anti-cofilin antibodies (1:100 dilution, #SC365882 and #SC376476, Santa, USA).

Techniques: Phospho-proteomics, Activation Assay, Inhibition, Activity Assay, Western Blot, Flow Cytometry

Journal: Materials Today Bio

Article Title: Stiff matrix promotes lung cancer cell migration through down-regulating the Piezo1 channel expression to facilitate Ca 2+ -dependent filopodia formation

doi: 10.1016/j.mtbio.2026.102786

Figure Lengend Snippet: Schematic summary of the Piezo1/calcium/CaN-SSH/cofilin/filopodia formation pathway for substrate stiffness-induced cancer cell migration. Stiff matrix downregulates the expression of Piezo1 channel, which limits the [Ca 2+ ] i rise and the activity of CaN-SSH. Consequently, cofilin is phosphorylated and inactivated and thereby loses its potential to bind to and sever actin filaments, facilitates filopodia formation, and thereby promote cancer cell migration.

Article Snippet: The following primary antibodies were used: rabbit anti-Piezo1 antibody (1:100 dilution, #DF12083, Affinity Biosciences, China) and rabbit anti-β-actin antibody (1:500 dilution, #GB11001-100, Servicebio, China), anti-p-cofilin and anti-cofilin antibodies (1:100 dilution, #SC365882 and #SC376476, Santa, USA).

Techniques: Migration, Expressing, Activity Assay

Journal: Journal of Nanobiotechnology

Article Title: f-SPION-mediated magnetic stimulation induces reparative Schwann cell reprogramming via cytoskeletal dynamics - gated activation of Piezo1

doi: 10.1186/s12951-026-04105-x

Figure Lengend Snippet: f-SPIONs-mediated magnetic actuation upregulates Piezo1 expression via activation of actin cytoskeleton dynamics. a RT-qPCR analysis of Piezo1 mRNA expression levels across different peripheral nerve tissues. b, c ( b ) RT-qPCR and ( c ) Western blot analyses of Piezo1 expression in Schwann cells from normal control, magnetic stimulation, f-SPIONs control, and magnetic field control groups. d Immunofluorescence imaging of F-actin and Piezo1 expression and subcellular distribution across different treatment groups: ( d 1 ) normal control, ( d 2 ) magnetic stimulation, ( d 3 ) f-SPIONs only, and ( d 4 ) magnetic field only groups. e Quantitative analysis of fluorescence intensity of F-actin and Piezo1 across different treatment groups. f Subcellular fluorescence distribution of F-actin and Piezo1 within soma and neurite compartments under magnetic stimulation. g, h Piezo1 expression levels measured by ( g ) RT-qPCR and ( h ) Western blot in magnetic stimulation, Cyto D, magnetic + Cyto D, and magnetic + Anisomycin groups. i Immunofluorescence imaging of F-actin and Piezo1 expression in SCs treated with ( i 1 ) magnetic stimulation, ( i 2 ) Cyto D, ( i 3 ) magnetic stimulation + Cyto D, and ( i 4 ) magnetic stimulation + Anisomycin. j Quantitative comparison of F-actin and Piezo1 fluorescence intensity across different treatment groups. Data are representative of three independent replicates. “n” indicates the total number of cells measured. Statistical significance: * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The membranes were subsequently incubated overnight at 4 °C with specific primary antibodies: rabbit polyclonal anti-Piezo1 antibody (1:1000, Alomone Labs, Cat. #APC-087, Jerusalem, Israel), rabbit polyclonal anti-Piezo1 antibody (1:1000, Thermo Fisher Scientific, Cat. #PA5-116998, Waltham, MA, USA), rabbit monoclonal anti-NCAM antibody (neural cell adhesion molecule, 1:1000, Abcam, Cat. #ab220360, Cambridge, UK), rabbit monoclonal anti-integrin β1 antibody (1:1000, Abcam, Cat. #ab179471, Cambridge, UK), rabbit monoclonal anti-c-Jun antibody (1:1000, Abcam, Cat. #ab40766, Cambridge, UK), rabbit monoclonal anti-STAT3 antibody (1:1000, Abcam, Cat. #ab68153, Cambridge, UK), and mouse monoclonal anti-GAPDH antibody (1:1000, Proteintech, Cat. #60004-1-Ig, Wuhan, China).

Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Western Blot, Control, Immunofluorescence, Imaging, Fluorescence, Comparison

Journal: Journal of Nanobiotechnology

Article Title: f-SPION-mediated magnetic stimulation induces reparative Schwann cell reprogramming via cytoskeletal dynamics - gated activation of Piezo1

doi: 10.1186/s12951-026-04105-x

Figure Lengend Snippet: f-SPION-mediated magnetic actuation activates Ca 2 ⁺ dynamics in Schwann cells via Piezo1 ion channel signaling. a 1 - d 1 Representative Ca 2 ⁺ fluorescence images of SCs in the ( a 1 ) normal control, ( b 1 ) magnetic stimulation, ( c1 ) f-SPIONs control, and ( d 1 ) magnetic field control groups. a 2 -d 2 Corresponding Ca 2 ⁺ signal trajectories from the four groups, showing temporal changes in intracellular calcium. e, f Quantification of ( e ) relative fluorescence intensity change (ΔF/F₀) and ( f ) peak fluorescence intensity (F max /F 0 ) of Ca 2 ⁺ signaling in different treatment groups. g 1 -j 1 Representative Ca 2 ⁺ fluorescence images in ( g 1 ) magnetic stimulation, ( h 1 ) Cyto D, ( i 1 ) magnetic stimulation + Cyto D, and (j 1 ) magnetic stimulation + Anisomycin groups. g 2 -j 2 Corresponding Ca 2 ⁺ signal trajectories showing real-time intracellular Ca 2 ⁺ fluctuations under the above treatments. k, l Quantification of ( k ) ΔF/F₀ and ( l ) F max /F 0 in different cytoskeleton intervention groups. m 1 -p 1 Representative Ca 2 ⁺ fluorescence imaging of SCs in ( m 1 ) magnetic stimulation, ( n 1 ) GsMTx4 (Piezo1 inhibitor), ( o 1 ) magnetic stimulation + GsMTx4, and ( p 1 ) Yoda1 (Piezo1 agonist) groups. m 2 -p 2 Corresponding Ca 2 ⁺ signal trajectories under the above intervention groups. q, r Quantification of ( q ) ΔF/F₀ and ( r ) F max /F 0 across Piezo1-specific intervention groups. Each experiment was independently repeated three times. “n” indicates the total number of cells measured. Baseline fluorescence intensity (F₀) was normalized to the first 50 s of imaging. GMF exposure was initiated after 50 s, and Ca 2 ⁺ time-lapse imaging continued for a total of 650 s. Statistical significance: * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The membranes were subsequently incubated overnight at 4 °C with specific primary antibodies: rabbit polyclonal anti-Piezo1 antibody (1:1000, Alomone Labs, Cat. #APC-087, Jerusalem, Israel), rabbit polyclonal anti-Piezo1 antibody (1:1000, Thermo Fisher Scientific, Cat. #PA5-116998, Waltham, MA, USA), rabbit monoclonal anti-NCAM antibody (neural cell adhesion molecule, 1:1000, Abcam, Cat. #ab220360, Cambridge, UK), rabbit monoclonal anti-integrin β1 antibody (1:1000, Abcam, Cat. #ab179471, Cambridge, UK), rabbit monoclonal anti-c-Jun antibody (1:1000, Abcam, Cat. #ab40766, Cambridge, UK), rabbit monoclonal anti-STAT3 antibody (1:1000, Abcam, Cat. #ab68153, Cambridge, UK), and mouse monoclonal anti-GAPDH antibody (1:1000, Proteintech, Cat. #60004-1-Ig, Wuhan, China).

Techniques: Fluorescence, Control, Imaging

Journal: Journal of Nanobiotechnology

Article Title: f-SPION-mediated magnetic stimulation induces reparative Schwann cell reprogramming via cytoskeletal dynamics - gated activation of Piezo1

doi: 10.1186/s12951-026-04105-x

Figure Lengend Snippet: Magnetically triggered Ca 2 ⁺ dynamics induce Schwann cell reprogramming toward a reparative phenotype. a Schematic diagram of the experimental workflow investigating the regulatory effect of magnetically triggered Ca 2 ⁺ dynamics activation on the reparative phenotype of SCs. b Live-cell Ca 2 ⁺ fluorescence imaging showing the effects of different treatments on SC structural remodeling and morphological polarization: ( b 1 ) magnetic stimulation, ( b 2 ) GsMTx4 (Piezo1 inhibitor), ( b 3 ) magnetic stimulation + GsMTx4, and ( b 4 ) Yoda1 (Piezo1 agonist). c , d Quantification of ( c ) cell length and ( d ) Oi across Piezo1-specific intervention groups. e , f Expression of the transcription factor c-Jun measured by ( e ) RT-qPCR and ( f ) Western blot. g , h STAT3 expression levels evaluated using ( g ) RT-qPCR and ( h ) Western blot. i , j NCAM expression assessed by ( i ) RT-qPCR and ( j ) Western blot. k , l Expression of integrin β1 determined via ( k ) RT-qPCR and ( l ) Western blot. Each experiment was independently repeated three times. “n” indicates the total number of cells measured. Statistical significance: n.s., not significant; * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The membranes were subsequently incubated overnight at 4 °C with specific primary antibodies: rabbit polyclonal anti-Piezo1 antibody (1:1000, Alomone Labs, Cat. #APC-087, Jerusalem, Israel), rabbit polyclonal anti-Piezo1 antibody (1:1000, Thermo Fisher Scientific, Cat. #PA5-116998, Waltham, MA, USA), rabbit monoclonal anti-NCAM antibody (neural cell adhesion molecule, 1:1000, Abcam, Cat. #ab220360, Cambridge, UK), rabbit monoclonal anti-integrin β1 antibody (1:1000, Abcam, Cat. #ab179471, Cambridge, UK), rabbit monoclonal anti-c-Jun antibody (1:1000, Abcam, Cat. #ab40766, Cambridge, UK), rabbit monoclonal anti-STAT3 antibody (1:1000, Abcam, Cat. #ab68153, Cambridge, UK), and mouse monoclonal anti-GAPDH antibody (1:1000, Proteintech, Cat. #60004-1-Ig, Wuhan, China).

Techniques: Activation Assay, Fluorescence, Imaging, Expressing, Quantitative RT-PCR, Western Blot

Journal: Nature Materials

Article Title: Long-range chemical signalling in vivo is regulated by mechanical signals

doi: 10.1038/s41563-025-02463-9

Figure Lengend Snippet: a , Schematic of the lateral view of a stage 40 Xenopus brain with orientation guides for rostral–caudal (R, C) and dorsal–ventral (D, V) axes. RGC axons grow from the optic chiasm towards the optic tectum along a stereotypic path (the optic tract), turning caudally at the mid-diencephalon (marked by an asterisk). b , Schematic cross-section of a Xenopus brain and retinae when Piezo1 is downregulated unilaterally in the nervous system. RGC axons cross the midline at the optic chiasm and grow across the contralateral brain surface. Normal Piezo1 levels are indicated in blue while Piezo1 depletion is shown in red. Piezo1-depleted axons grow across brain tissue with normal Piezo1 levels and vice versa. c – f , Images of RGC axon growth in vivo in control ( c ), Piezo1-depleted axons ( d ), Piezo1-depleted surrounding brain tissue (parenchyma) ( e ), and both axons and brain tissue depleted of Piezo1 ( f ). Scale bars, 50 μm. g , Optic tract (OT) elongation. Schematic representation of the fitted ellipse used to determine optic tract elongation, expressed as the ratio of long to short axes. Quantification for the indicated conditions (Kruskal–Wallis test, P < 0.0001, followed by Dunn’s post hoc test; adjusted P -values indicated). Each point represents a brain. Boxes show median, first and third quartiles; whiskers show the spread of data; ‘+’ indicate outliers. N denotes the number of animals. h , Scoring of brains displaying aberrant phenotypes. Quantification of embryos showing normal, stalling or misprojection defects after Piezo1 knockdown in axons, brain tissue or both (two-tailed chi-squared test, P = 1.014 × 10 −8 , followed by Fisher’s exact post hoc tests; number of animals indicated in parentheses) (Extended Data Fig. ). Data are pooled from a minimum of three independent experiments. chi, chiasm; di, diencephalon; KD, knockdown; tec, tectum; tel, telencephalon.

Article Snippet: Samples were then incubated with rabbit anti-Piezo1 antibody (Proteintech, 28511-1-AP, 1:10 dilution in blocking solution) at 4 °C overnight.

Techniques: In Vivo, Control, Knockdown, Two Tailed Test

Journal: Nature Materials

Article Title: Long-range chemical signalling in vivo is regulated by mechanical signals

doi: 10.1038/s41563-025-02463-9

Figure Lengend Snippet: a , Schematic. At the 4-cell stage, translation-blocking morpholinos were injected into one of the two dorsal (usually less pigmented) blastomeres, which contribute to the formation of the nervous system. Scale bar: 600 µm. b-d , Immunofluorescence images of Xenopus laevis brains at stage 40. The dashed white outlines depict the boundaries of the brain tissue. Scale bars: 100 µm. b-c , Piezo1 expression patterns in neuroepithelium (lateral view) arising from the b , uninjected blastomere and c , the blastomere injected with translation-blocking Piezo1 morpholino, resulting in Piezo1 knockdown (KD). d , Dorsal view of Piezo1 expression in the developing neuroepithelium showing both the injected (Piezo1 KD) and the uninjected (Control) hemispheres. The dashed yellow line represents the midline. e , Piezo1 morpholino injection into one dorsal blastomere led to a significant decrease in Piezo1 expression in one half of the developing neuroepithelium. The graph represents the paired (Control vs Piezo1 KD) mean intensity values of the developing telencephalon. Each point represents an embryo ( N = 4). Data was assessed with a two tailed ratio paired t-test; p -value indicated in the figure. KD: knockdown.

Article Snippet: Samples were then incubated with rabbit anti-Piezo1 antibody (Proteintech, 28511-1-AP, 1:10 dilution in blocking solution) at 4 °C overnight.

Techniques: Blocking Assay, Injection, Immunofluorescence, Expressing, Knockdown, Control, Two Tailed Test

Journal: Nature Materials

Article Title: Long-range chemical signalling in vivo is regulated by mechanical signals

doi: 10.1038/s41563-025-02463-9

Figure Lengend Snippet: a , Schematic representation of the expression pattern of the diffusive chemical guidance cue, Slit1. b , c , Representative HCR-FISH images of Slit1 expression in control ( b ) and Piezo1 knockdown ( c ) brains. d , Quantification of Slit1 mRNA expression (two-tailed unpaired t -test with Welch’s correction; P value indicated). e , Schematic of the Sema3A expression pattern. f , g , Representative HCR-FISH images of Sema3A expression in control ( f ) and Piezo1 knockdown ( g ) brains. h , Quantification of normalized Sema3A expression (two-tailed unpaired t-test with Welch’s correction; P value indicated). i , j , In situ hybridization of Sema3A mRNA expression in control ( i ) and Piezo1 knockdown ( j ) brains. k , Western blot of Piezo1, Sema3A and β-actin protein expression in control and Piezo1-depleted brains. l , Western blot quantification ( N = 4, normalized to total protein, each point indicates the mean value of a biological replicate; two-tailed ratio paired t -test; P value indicated). Each point in d , h represents an embryo; bars indicate lower quartile, median and upper quartiles. N , number of animals. Scale bars, 100 μm. ISH, in situ hybridization.

Article Snippet: Samples were then incubated with rabbit anti-Piezo1 antibody (Proteintech, 28511-1-AP, 1:10 dilution in blocking solution) at 4 °C overnight.

Techniques: Expressing, Control, Knockdown, Two Tailed Test, In Situ Hybridization, Western Blot

Journal: Nature Materials

Article Title: Long-range chemical signalling in vivo is regulated by mechanical signals

doi: 10.1038/s41563-025-02463-9

Figure Lengend Snippet: Downregulating Piezo1 led to a significant decrease in the expression of Piezo1 protein a , but not of β-actin b . Representative Western blot images are shown in Fig. . Data were normalised by the total protein concentration. Each point represents the mean of a biological replicate (N = 4; two-tailed ratio paired t-test; p-values indicated). KD: knockdown.

Article Snippet: Samples were then incubated with rabbit anti-Piezo1 antibody (Proteintech, 28511-1-AP, 1:10 dilution in blocking solution) at 4 °C overnight.

Techniques: Expressing, Western Blot, Protein Concentration, Two Tailed Test, Knockdown

Journal: Nature Materials

Article Title: Long-range chemical signalling in vivo is regulated by mechanical signals

doi: 10.1038/s41563-025-02463-9

Figure Lengend Snippet: a , Schematic of the experimental set-up for in vivo brain-stiffness mapping. b , Xenopus brain schematics. Dashed rectangle indicates the mapped region; lower left (LL) and upper right (UR) corners of stiffness maps ( c , f ) are indicated, colours indicate areas selected for regional analysis ( g , h ). c , AFM-based stiffness maps (colour maps) encoding the apparent elastic modulus, K , a measure of tissue stiffness, assessed at an indentation force F = 10 nN for control or Piezo1 downregulation exclusively in axons, in the surrounding brain tissue, or both. d , Quantification of AFM measurements of stiffness in different conditions (Kruskal–Wallis test, P < 0.0001; Dunn’s post hoc tests, adjusted P values indicated). e , Exposed brain of stage 40 Xenopus embryo electroporated with fluorescein-tagged morpholinos to visualize electroporated regions. Dashed lines: brain outline (white), electroporated region (blue) and AFM cantilever (black). Scale bar, 250 µm. f – h , Downregulating Sema3A and assessing tissue stiffness: AFM-based stiffness maps for control or Sema3A morpholino-electroporated brains ( f ); quantification of AFM stiffness measurements in Sema3A-producing regions ( g ) and adjacent non-Sema3A-producing regions ( h ) (Wilcoxon rank-sum test, P values indicated). Violin plots in d , g , h display the data distribution, overlayed with individual measurements shown as scattered points; means and medians are indicated as yellow crosses and white squares, respectively. Scale bars, 100 μm. N , number of animals; n , number of measurements.

Article Snippet: Samples were then incubated with rabbit anti-Piezo1 antibody (Proteintech, 28511-1-AP, 1:10 dilution in blocking solution) at 4 °C overnight.

Techniques: In Vivo, Control

Journal: Nature Materials

Article Title: Long-range chemical signalling in vivo is regulated by mechanical signals

doi: 10.1038/s41563-025-02463-9

Figure Lengend Snippet: a, b , Heatmaps of AFM measurements for stage 31 a , SCR (control) and b , Piezo1 knockdown (KD) embryo skin tissue, overlaid on a fluorescence image of the GFP-containing injected constructs. Heatmaps are scaled to an apparent elastic modulus of 170 Pa; scale bar is 200 μm. c , Median skin tissue stiffness per animal in both conditions. Each dot corresponds to one animal; boxes span the first and third quartiles with a line at the median. The whiskers span the range of the data. Both groups were compared with a nested t-test, where AFM measurements were nested under animals, and animals under condition. N = 13 animals per condition; n = 3 - 23 stiffness measurements per animal.

Article Snippet: Samples were then incubated with rabbit anti-Piezo1 antibody (Proteintech, 28511-1-AP, 1:10 dilution in blocking solution) at 4 °C overnight.

Techniques: Control, Knockdown, Fluorescence, Injection, Construct

Journal: Nature Materials

Article Title: Long-range chemical signalling in vivo is regulated by mechanical signals

doi: 10.1038/s41563-025-02463-9

Figure Lengend Snippet: a , b , Local cell body densities. a , Representative images of nuclei (DAPI, magenta) in brain tissue rostral (R) and caudal (C) to the optic tract (DiI, yellow) in stage 40 Xenopus embryos. Scale bars, 50 μm. b , Relative nuclear area rostral and caudal to the optic tract (two independent experiments; one-way ANOVA). c , Western blot of NCAM1, N-cadherin, acetylated α-tubulin and total α-tubulin in control and Piezo1-depleted brains. d , Western blot quantification ( N = 4) of relative proportion of acetylated α tubulin (ratio of acetylated α-tubulin/total α-tubulin). e , Quantification of the reduced apparent elastic modulus K of dissociated brain cells (two-tailed Mann–Whitney test). Each point represents a single cell. f , g , Western blot quantification ( N = 3; normalized to total protein) of NCAM1 (250 kDa) ( f ) and N-cadherin ( g ). h , AFM-based stiffness maps overlaid on bright-field images of control and NCAM1- and N-cadherin-depleted brains. Scale bar, 100 µm. i , Tissue stiffness quantification (nested t -test). Each point represents the median K of an embryo. j , Schematic illustrating the mechanism linking Piezo1 to tissue stiffness: Piezo1 regulates major cell–cell adhesion proteins (N-cadherin, NCAM1), which in turn regulate tissue stiffness. k , Western blot of Sema3A protein expression in control and NCAM1 and N-cadherin-depleted brains. l , Quantification of Sema3A protein expression ( N = 3; normalized to total protein). d , f , g , l , show two-tailed ratio paired t -tests; P- values are indicated. In b , e , i boxes show first and third quartiles with median lines; whiskers show the spread of data. Double KD, optic tract and brain tissue depleted of Piezo1.

Article Snippet: Samples were then incubated with rabbit anti-Piezo1 antibody (Proteintech, 28511-1-AP, 1:10 dilution in blocking solution) at 4 °C overnight.

Techniques: Western Blot, Control, Two Tailed Test, MANN-WHITNEY, Single Cell, Expressing

Journal: Nature Materials

Article Title: Long-range chemical signalling in vivo is regulated by mechanical signals

doi: 10.1038/s41563-025-02463-9

Figure Lengend Snippet: a , Schematic of the experimental set-up for locally increasing tissue stiffness in wild-type brains in vivo. The hypothalamus was compression-stiffened with an AFM probe for >6 h. b , c , Representative Sema3A HCR-FISH images of control ( b ) and compression-stiffened ( c ) brains. Insets: regions selected for analysis. d , Ratio of the total area covered by signal in the compression-stiffened (CS) region to the mean area covered by the signal in background (BG) regions were analysed (unpaired t -test with Welch’s correction; P value indicated). e , Schematic of the experimental set-up for locally increasing tissue stiffness in Piezo1 knockdown brains in vivo. The telencephalic region of the brain was compression-stiffened as in a . f , g , Representative Sema3A HCR-FISH images in control ( f ) and compression-stiffened ( g ) Piezo1 knockdown brains. Insets: regions selected for analysis. h , Ratio of the total area covered by signal in the CS region to the mean area covered by the signal in BG regions were analysed (Kruskal–Wallis test, P < 0.0001; Dunn’s post hoc test for multiple comparison; P values indicated). In d , h , each point represents an embryo; lower quartile, median and upper quartile are indicated by bars. Scale bars, 100 μm (whole brain), 20 μm (insets). SCR, scrambled control morpholino.

Article Snippet: Samples were then incubated with rabbit anti-Piezo1 antibody (Proteintech, 28511-1-AP, 1:10 dilution in blocking solution) at 4 °C overnight.

Techniques: In Vivo, Control, Knockdown, Comparison