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