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: In some cases, cells were cultured in medium containing 2 μM Yoda1 (#M9372, AbMole, USA) to activate the Piezo1 channel, 2.5 μM GsMTx4 (#HY-P1410, MedChemExpress, USA) to block the Piezo1 channel, 25 μM BAPTA-AM (#M4973, AbMole, USA) to chelate intracellular Ca 2+ or adding 2 mM CaCl 2 to observe extracellular Ca 2+ influx, 4 μM cyclosporine A (CsA) (#M1831, AbMole, USA) to inhibit CaN, 0.5 μM dasatinib (Das, #9052S, Cell signaling technology) to inhibit YAP nuclear translocation, as well as 10 μM PF-573228 (PF, #HY-10461, MCE) to inhibit focal adhesion kinase (FAK) activation, respectively.

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: In some cases, cells were cultured in medium containing 2 μM Yoda1 (#M9372, AbMole, USA) to activate the Piezo1 channel, 2.5 μM GsMTx4 (#HY-P1410, MedChemExpress, USA) to block the Piezo1 channel, 25 μM BAPTA-AM (#M4973, AbMole, USA) to chelate intracellular Ca 2+ or adding 2 mM CaCl 2 to observe extracellular Ca 2+ influx, 4 μM cyclosporine A (CsA) (#M1831, AbMole, USA) to inhibit CaN, 0.5 μM dasatinib (Das, #9052S, Cell signaling technology) to inhibit YAP nuclear translocation, as well as 10 μM PF-573228 (PF, #HY-10461, MCE) to inhibit focal adhesion kinase (FAK) activation, respectively.

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: In some cases, cells were cultured in medium containing 2 μM Yoda1 (#M9372, AbMole, USA) to activate the Piezo1 channel, 2.5 μM GsMTx4 (#HY-P1410, MedChemExpress, USA) to block the Piezo1 channel, 25 μM BAPTA-AM (#M4973, AbMole, USA) to chelate intracellular Ca 2+ or adding 2 mM CaCl 2 to observe extracellular Ca 2+ influx, 4 μM cyclosporine A (CsA) (#M1831, AbMole, USA) to inhibit CaN, 0.5 μM dasatinib (Das, #9052S, Cell signaling technology) to inhibit YAP nuclear translocation, as well as 10 μM PF-573228 (PF, #HY-10461, MCE) to inhibit focal adhesion kinase (FAK) activation, respectively.

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: In some cases, cells were cultured in medium containing 2 μM Yoda1 (#M9372, AbMole, USA) to activate the Piezo1 channel, 2.5 μM GsMTx4 (#HY-P1410, MedChemExpress, USA) to block the Piezo1 channel, 25 μM BAPTA-AM (#M4973, AbMole, USA) to chelate intracellular Ca 2+ or adding 2 mM CaCl 2 to observe extracellular Ca 2+ influx, 4 μM cyclosporine A (CsA) (#M1831, AbMole, USA) to inhibit CaN, 0.5 μM dasatinib (Das, #9052S, Cell signaling technology) to inhibit YAP nuclear translocation, as well as 10 μM PF-573228 (PF, #HY-10461, MCE) to inhibit focal adhesion kinase (FAK) activation, respectively.

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: In some cases, cells were cultured in medium containing 2 μM Yoda1 (#M9372, AbMole, USA) to activate the Piezo1 channel, 2.5 μM GsMTx4 (#HY-P1410, MedChemExpress, USA) to block the Piezo1 channel, 25 μM BAPTA-AM (#M4973, AbMole, USA) to chelate intracellular Ca 2+ or adding 2 mM CaCl 2 to observe extracellular Ca 2+ influx, 4 μM cyclosporine A (CsA) (#M1831, AbMole, USA) to inhibit CaN, 0.5 μM dasatinib (Das, #9052S, Cell signaling technology) to inhibit YAP nuclear translocation, as well as 10 μM PF-573228 (PF, #HY-10461, MCE) to inhibit focal adhesion kinase (FAK) activation, respectively.

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: In some cases, cells were cultured in medium containing 2 μM Yoda1 (#M9372, AbMole, USA) to activate the Piezo1 channel, 2.5 μM GsMTx4 (#HY-P1410, MedChemExpress, USA) to block the Piezo1 channel, 25 μM BAPTA-AM (#M4973, AbMole, USA) to chelate intracellular Ca 2+ or adding 2 mM CaCl 2 to observe extracellular Ca 2+ influx, 4 μM cyclosporine A (CsA) (#M1831, AbMole, USA) to inhibit CaN, 0.5 μM dasatinib (Das, #9052S, Cell signaling technology) to inhibit YAP nuclear translocation, as well as 10 μM PF-573228 (PF, #HY-10461, MCE) to inhibit focal adhesion kinase (FAK) activation, respectively.

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