human a549  (ATCC)

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: MagGO induces tumor cell death via frequency- and mode-dependent mechanical disruption (A) TEM images of MagGO synthesized under MF. The dashed white line represents the contour edge of GO. Scale bars, 200 nm. (B) M-H curve of GO, MNP, and MagGO. (C) AFM image of MagGO and the height of GO in MagGO. Scale bars, 500 nm. (D, F, and H) Cell viability of U87 (D), MDA-MB-231 (F), and A549 (H) cells treated with MNP and MagGO under RMF and 3D MF (RMF combined with OMF stimulation) at 5 Hz. The applied field strength is 75 mT. The data were presented as the mean ± SD. (E, G, and I) Cell viability of U87 (E), MDA-MB-231 (G), and A549 (I) cells treated with MNP and MagGO under 3D MF of different frequencies. The applied field strength is 75 mT. The data were presented as the mean ± SD.

Article Snippet: Human: A549 , ATCC , Cat#CCL-185.

Techniques: Disruption, Synthesized

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: MagGO induces lysosomal disruption (A) CLSM images of MNPs and MagGO in U87 cells. Lysosomes were stained with LysoTracker red (red), and the MNPs and MagGO were labeled with FITC (green). Scale bars, 15 μm. (B and C) Intensity profiles (white dashed line) of signals from lysosome and MNPs/MagGO fluorescent channels in (A). (D) CLSM images of MNPs and MagGO in MDA-MB-231 cells. Lysosomes were stained with LysoTracker red (red), and the MNPs and MagGO were labeled with FITC (green). Scale bars, 15 μm. (E and F) Intensity profiles (white dashed line) of signals from lysosomes and MNPs/MagGO fluorescent channels in (D). (G) CLSM images of MNPs and MagGO in A549 cells. Lysosomes were stained with LysoTracker red (red), and the MNPs and MagGO were labeled with FITC (green). Scale bars, 15 μm. (H and I) Intensity profiles (white dashed line) of signals from lysosomes and MNPs/MagGO fluorescent channels in (G). (J) CLSM images of U87 cells transfected with the EGFP-Gal3 plasmid after MagGO treatment under 3D MF. The applied field strength is 75 mT. The duration of magnetic field application is 30 min. Scale bars, 15 μm. (K) Counts of Gal3 puncta per U87 cell ( n = 10). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (L) Counts of Gal3 puncta per MDA-MB-231 cell ( n = 10). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (M) Counts of Gal3 puncta per A549 cell ( n = 10). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (N) Bio-TEM of lysosomal membrane morphology after mechanoporation for MagGO and MagGO+3D MF. The applied field strength is 75 mT. The duration of magnetic field application is 30 min. Scale bars, 1 μm. The dark blue arrow indicates the site of LMP, while the length of the blue arrow represents the size of the lysosomal membrane “wound”.

Article Snippet: Human: A549 , ATCC , Cat#CCL-185.

Techniques: Disruption, Staining, Labeling, Transfection, Plasmid Preparation, Membrane

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: MagGO primarily induces pyroptosis as the mode of cell death (A–C) CLSM images of CTSB release of U87 (A), MDA-MB-231 (B), and A549 (C) cells after MagGO treatment under 3D MF. The applied field strength is 75 mT. The duration of magnetic field application is 30 min. Scale bars, 20 μm. (D–F) The cell viabilities of U87 (D), MDA-MB-231 (E), and A549 (F) cells treated with MagGO were assessed after pre-treatment with z-VAD-FMK (10 μM), Necrostatin-1 (10 μM), 3-Methyladenine (10 μM), Ferrostatin-1 (2 μM), and MCC950 (10 nM) for 4 h, followed by exposure to 3D MF at 5 Hz for 30 min. The applied field strength was 75 mT. The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post hoc test. (G and H) Quantification of IL-1β (G) and IL-18 (H) release from U87 cells for control, MagGO, and MagGO+3D MF ( n = 3). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (I and J) Western blot analysis of Casp-1 (I) and GSDMD (J) in U87 cells for control, MagGO, and MagGO+3D MF.

Article Snippet: Human: A549 , ATCC , Cat#CCL-185.

Techniques: Control, Western Blot

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: Broad antitumor activity of MagGO across multiple tumor models (A) Schematic illustrations of in vivo anticancer therapy for MDA-MB-231 and A549 tumors. The applied field strength is 75 mT. The applied field frequency is 5 Hz. The duration of magnetic field application is 30 min. (B) The MDA-MB-231 tumor volume comparison of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF for 14 days ( n = 5). (C) The MDA-MB-231 tumor images of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). (D) The MDA-MB-231 tumor weight of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post hoc test. (E) The MDA-MB-231 tumor volume of each mouse in the control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF groups for 14 days. (F) The A549 tumor volume comparison of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF for 14 days ( n = 5). (G) The A549 tumor images of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). (H) The A549 tumor weight of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post hoc test. (I) The A549 tumor volume of each mouse in the control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF groups for 14 days.

Article Snippet: Human: A549 , ATCC , Cat#CCL-185.

Techniques: Activity Assay, In Vivo, Comparison, Control

Journal: Redox Biology

Article Title: Amino acid restriction sensitizes lung cancer cells to ferroptosis via GCN2-dependent activation of the integrated stress response

doi: 10.1016/j.redox.2025.103988

Figure Lengend Snippet: Increased mitochondrial respiration renders F12-cultured cells BSO-sensitive. ( A, B ) Oxygen consumption rate (left) in F12 or F12AA-cultured A549 (A) or H838 (B) cells treated with 0.5 μM oligomycin (Oligo), 1 μM FCCP, and 0.5 μM rotenone (Rot), as indicated. Graphs (right) showing parameter data extracted from the oxygen consumption rate and extracellular acidification rate (n = 14 in A549 cells; n = 10 in F12 and 4 in F12AA-cultured H838 cells). ( C , D ) Oxygen consumption rate (left) in F12-cultured A549 (C) or H838 (D) cells transfected with control siRNA or siRNA targeting ATF4 mRNA and assayed as in (A, B). Graphs (right) show extracted parameters (n = 15 for A549; n = 21 for ATF4 siRNA and n = 13 for control siRNA in H838). ( E ) MitoSox fluorescence images of A549 (left) and H838 (right) cells cultured in F12 or F12AA medium. Cells were treated with 30 μM BSO for 40 h (A549) or 48 h (H838). Graphs show IncuCyte-based fluorescence quantification over time. Scale bar, 100 μm. ( F ) MitoSox fluorescence images of A549 cells cultured in F12 or F12AA medium and treated for 24h with 50 μM BSO, 50 μM BSO + 20 μM mito-TEMPO (MT), or control. MitoSox was added during the last 90 min of treatment; cells were then washed and quantified by live imaging. The graph shows IncuCyte-based fluorescence quantification. Scale bar, 100 μm; n = 8–10. ( G ) Confocal microscopy images of F12-cultured A549 cells treated with 100 μM BSO for 24h showing reduced (pink) and oxidized (green) BODIPY-C11 in combination with mitotracker deep red (red). Corresponding graphs show pixel-wise colocalization (Mander's coefficient) of oxidized BODIPY-C11 and mitotracker deep red in F12-cultured A549 (left) or H838 (right) cells treated with 100 μM BSO for 24 h or controls. n = 6–9 visual fields in A549 cells and 48 visual fields in H838 cells. Scale bar, 10 μm. ( H ) Oxidized BODIPY-C11 fluorescence images of A549 cells cultured in F12 medium and treated for 12 h with 100 μM BSO, 100 nM rotenone, 100 nM oligomycin, or combinations of BSO + rotenone or BSO + oligomycin, and controls. Graph shows IncuCyte-based fluorescence quantification. Scale bar, 50 μm. ( I ) Oxidized BODIPY-C11 fluorescence images of A549 cells cultured in F12 medium and treated for 24 h with 100 μM BSO, BSO + 20 μM mito-TEMPO (MT), or control. Graph shows IncuCyte-based fluorescence quantification. Scale bar, 100 μm. ( J ) Oxidized MitoPerOx fluorescence images of A549 (left) and H838 (right) cells cultured in F12 or F12AA medium. Cells were treated with 30 μM BSO or control for 48 h. Graphs show IncuCyte-based fluorescence quantification over time. Scale bar, 100 μm. ( K) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 100 nM oligomycin, 100 nM rotenone, 20 μM mito-TEMPO + rotenone, or mito-TEMPO + oligomycin, or control for 72 h. ( L ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 100 nM oligomycin, 100 nM rotenone, 5 μM ferrostatin-1 (FER) + rotenone, 5 μM liproxstatin-1 (LIP) + rotenone, ferrostatin-1 + oligomycin, or liproxstatin-1 + oligomycin, or control for 72 h. ( M ) Dose response curves of F12-cultured A549 cells treated with BSO in combination with 0.5 μM FCCP, or control for 72 h. ( N ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 25 or 50 μM mito-TEMPO, or control for 72 h. Dose response curves were normalized against the mean of the untreated samples for each condition. n = 3 replicates for all datapoints unless otherwise indicated. Error bars show SEM. ∗∗∗∗P < 0.0001, ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05.

Article Snippet: Cas9-expressing A549 cells were obtained from GeneCopoeia (SL-504; GeneCopoeia, Inc., Rockville, MD) and the mouse KP cell line was obtained from V. Sayin and is described [ ].

Techniques: Cell Culture, Transfection, Control, Fluorescence, Imaging, Confocal Microscopy

Journal: Redox Biology

Article Title: Amino acid restriction sensitizes lung cancer cells to ferroptosis via GCN2-dependent activation of the integrated stress response

doi: 10.1016/j.redox.2025.103988

Figure Lengend Snippet: Culture in F12 medium sensitizes lung cancer cells to BSO. ( A ) Crystal violet staining of A549 cells that were cultured in RPMI or F12 medium, after treatment with 100 μM BSO or vehicle (Ctrl) for 72 h. ( B ) BSO dose response curves for A549, H838, H1299, H23, and H460 cells cultured in RPMI or F12 medium for 72 h. ( C ) Crystal violet staining and quantification of mouse KP cells that were cultured in RPMI or F12 medium in the presence of BSO at the indicated concentrations for 72 h. ( D ) Dose response curves for A549 cells cultured in RPMI or F12 medium and treated with erastin, RSL3, or auranofin for 72 h. Dose response curves were normalized against the mean of the untreated samples for each condition. n = 3 replicates for all datapoints, error bars show SEM.

Article Snippet: Cas9-expressing A549 cells were obtained from GeneCopoeia (SL-504; GeneCopoeia, Inc., Rockville, MD) and the mouse KP cell line was obtained from V. Sayin and is described [ ].

Techniques: Staining, Cell Culture

Journal: Redox Biology

Article Title: Amino acid restriction sensitizes lung cancer cells to ferroptosis via GCN2-dependent activation of the integrated stress response

doi: 10.1016/j.redox.2025.103988

Figure Lengend Snippet: F12 medium sensitizes lung cancer cells to iron-dependent lipid peroxidation . ( A, B ) Quantification by flow cytometry of oxidized BODIPY-C11 fluorescence in A549 (A) or H838 (B) cells that were cultured in RPMI or F12 medium and treated with 100 μM BSO or vehicle for 24 h. ( C ) Dose response curves for F12-cultured A549 and H838 cells treated with BSO in combination with 5 μM liproxstatin-1 (LIP-1), 5 μM ferrostatin-1 (FER-1) or 50 μM α-tocopherol (α-toco) for 72 h. ( D ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 5 μM deferoxamine (DFO) for A549 cells and 9 μM for H838 cells for 72 h. ( E ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 5 μM necrostatin-1 (NEC-1) or 10 μM ZVAD-FMK (ZVAD) for 72 h. ( F ) Dose response curves for F12-cultured A549 or H838 cells treated with BSO in combination with 4 μg/mL certolizumab (CER), 15 μM CU-CPT4a (C4a), 3 μM resatorvid (RST), or 50 μM necrostatin-1 (NEC-1) for 72 h (ND, not done). Dose response curves were normalized against the mean of the untreated samples for each condition. n = 3 replicates for all datapoints, error bars show SEM. ∗∗∗∗P < 0.0001.

Article Snippet: Cas9-expressing A549 cells were obtained from GeneCopoeia (SL-504; GeneCopoeia, Inc., Rockville, MD) and the mouse KP cell line was obtained from V. Sayin and is described [ ].

Techniques: Flow Cytometry, Fluorescence, Cell Culture

Journal: Redox Biology

Article Title: Amino acid restriction sensitizes lung cancer cells to ferroptosis via GCN2-dependent activation of the integrated stress response

doi: 10.1016/j.redox.2025.103988

Figure Lengend Snippet: The sensitizing effect of F12 medium is caused by lower amino acid content. ( A ) Concentrations of reduced glutathione in lysates of A549 cells cultured in RPMI or F12 medium and treated with the indicated concentrations of BSO for 24 h. ( B ) BSO dose response curves for A549 cells cultured in F12 or F12 medium supplemented with 65 mg/L cystine (F12 L-Cys) for 72 h. ( C ) BSO dose response curves for A549 and H838 cells cultured in F12 or F12AA medium, the latter with amino acid concentrations matching those of RPMI (see ), for 72 h. (D) BSO dose response curves for A549 cells cultured in RPMI or RPMIAA medium, the latter with amino acid concentrations matching those of F12 (see ), for 72 h. ( E, F ) GC/MS data for intracellular levels of serine, methionine, isoleucine, and leucine (E) or cysteine, glutamate, and glycine (F) in A549 cells at 1, 6, 24, and 48 h after switching from RPMI medium to F12 or F12AA medium. The cells were maintained in RPMI and then passaged into fresh RPMI for 24 h before being switched to F12, F12AA, or fresh RPMI. (G) GC/MS data showing uptake of serine, leucine, and isoleucine in A549 cells that were cultured in F12 or F12AA medium for 48 h. (H) Heatmap showing BSO dose responses of A549 cells cultured in F12 medium supplemented with the indicated amino acids at final concentrations matching the ones in RPMI (see ). (I) Concentrations of reduced glutathione in lysates of A549 and H838 cells cultured in F12 or F12AA medium and treated with the indicated concentrations of BSO for 24 h. Dose response curves were normalized against the mean of the untreated samples for each condition. n = 3 replicates for all datapoints, error bars show SEM. ∗∗P < 0.01, ∗P < 0.05.

Article Snippet: Cas9-expressing A549 cells were obtained from GeneCopoeia (SL-504; GeneCopoeia, Inc., Rockville, MD) and the mouse KP cell line was obtained from V. Sayin and is described [ ].

Techniques: Cell Culture, Gas Chromatography-Mass Spectrometry

Journal: Redox Biology

Article Title: Amino acid restriction sensitizes lung cancer cells to ferroptosis via GCN2-dependent activation of the integrated stress response

doi: 10.1016/j.redox.2025.103988

Figure Lengend Snippet: The integrated stress response pathway is activated in F12-cultured cells . ( A ) Western blotting of S6, p-S6, 4E-BP1, p-4E-BP1 in protein extracts of A549 cells cultured in F12, F12AA, or RPMI medium and treated with 100 μM BSO for 24 h. HSP90 was used as loading control. (B) Western blotting and quantification of p-S6 and p-4E-BP1 in protein extracts of A549 cells cultured in F12 medium and treated with 10 or 50 nM torin1 for 24 h. HSP90 was used as loading control. ( C ) Viability (luminescence) of F12-cultured A549 cells treated with 10 or 50 nM torin1 for 24 h. ( D ) BSO dose response curves for A549 cells cultured in F12 medium and treated with 10 nM torin1 or control for 72 h. The data were normalized against the mean of the untreated samples for each condition. (E) Schematic of the ISR pathway. ( F, G ) Western blotting and quantification of GCN2, p-GCN2, eIF2α, p-eIF2α, ATF4, and CHOP in protein extracts of A549 (F) or H838 (G) cells cultured in F12 or F12AA medium and treated with 100 μM BSO for 24 h. HSP90 was used as loading control. (H) Western blotting and quantification of p-GCN2 and ATF4 in protein extracts of A549 cells at 0, 6, 9, 12, 24, and 48 h after switching from F12AA medium to a pre-conditioned F12 medium. ( I ) Schematic model showing methionine abundance, estimated methionine abundance, ATF4 expression, and estimated ISR activity, as indicated. Data on methionine abundance were retrieved from E and ATF4 expression from H. Thresholds for mild and robust ISR activation are indicated by arrows. n = 3 replicates for all datapoints, error bars show SEM. ∗∗∗∗P < 0.0001, ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05.

Article Snippet: Cas9-expressing A549 cells were obtained from GeneCopoeia (SL-504; GeneCopoeia, Inc., Rockville, MD) and the mouse KP cell line was obtained from V. Sayin and is described [ ].

Techniques: Cell Culture, Western Blot, Control, Expressing, Activity Assay, Activation Assay

Journal: Redox Biology

Article Title: Amino acid restriction sensitizes lung cancer cells to ferroptosis via GCN2-dependent activation of the integrated stress response

doi: 10.1016/j.redox.2025.103988

Figure Lengend Snippet: Increased autophagy in F12-cultured cells does not influence BSO sensitivity . ( A, B ) Western blotting (A) and quantification (B) of LC3B–I and II expression in protein extracts of A549 cells cultured in F12 or F12AA medium and treated with 50 μM chloroquine (ChlQ) for the indicated time periods. HSP90 was used as loading control. (C) Western blotting and quantification of TFRC and Ferritin (heavy chain) in protein extracts of A549 cells cultured in F12 or F12AA medium. Tubulin was used as loading control. (D) Western blotting and quantification of LC3B-II in protein extracts of A549 cells cultured in F12 medium and treated with 20 nM torin1 for 24 h. Tubulin was used as loading control. (E) Dose response curves for F12-cultured A549 cells treated with BSO in combination with 20 nM torin1 or control for 72 h. ( F ) Western blotting and quantification of LC3B-II expression in protein extracts of A549 cells cultured in F12 medium and treated with 0, 0.6, or 5 mM 3-MA in the presence of 50 μM chloroquine (ChlQ) for 1 h. HSP90 was used as loading control. (G) Dose response curves for F12-cultured A549 cells treated with BSO in combination with 0, 0.6, 2.5, or 5 mM 3-MA for 72 h. Dose response curves were normalized against the mean of the untreated samples for each condition. n = 3 replicates for all datapoints, error bars show SEM. ∗∗P < 0.01, ∗P < 0.05.

Article Snippet: Cas9-expressing A549 cells were obtained from GeneCopoeia (SL-504; GeneCopoeia, Inc., Rockville, MD) and the mouse KP cell line was obtained from V. Sayin and is described [ ].

Techniques: Cell Culture, Western Blot, Expressing, Control

Journal: Redox Biology

Article Title: Amino acid restriction sensitizes lung cancer cells to ferroptosis via GCN2-dependent activation of the integrated stress response

doi: 10.1016/j.redox.2025.103988

Figure Lengend Snippet: Activation of the ISR pathway sensitizes lung cancer cells to BSO. ( A, B ) Western blotting of GCN2 in protein extracts of F12-cultured A549 cells (A) and GCN2, p-eIF2α, ATF4 and CHOP in protein extracts of F12-cultured H838 cells (B) that were transfected with Ctrl siRNA or siRNA targeting GCN2 mRNA. HSP90 was used as loading control. ( C ) BSO dose response curves for A549 (left) and H838 (right) cells cultured in F12 medium for 72 h and transfected with Ctrl siRNA or siRNA targeting GCN2 mRNA. ( D ) Western blotting and quantification of GADD34 in protein extracts of A549 cells that were cultured in F12 or F12AA medium and treated with 100 μM BSO for 24 h. Tubulin was used as loading control. ( E ) Western blotting of GADD34 in protein extracts of F12-cultured A549 cells that were transfected with Ctrl siRNA or siRNA targeting GADD34 mRNA. Tubulin was used as loading control. ( F ) Western blotting and quantification of p-eIF2α and CHOP in protein extracts of F12-cultured A549 cells that were transfected with Ctrl siRNA or siRNA targeting GADD34 mRNA. HSP90 was used as loading control. ( G ) BSO dose response curves for A549 cells cultured in F12 medium for 72 h and transfected with Ctrl siRNA or siRNA targeting GADD34 mRNA. ( H ) Western blotting of ATF4 in protein extracts of F12-cultured A549 (top) or H838 (bottom) cells that were transfected with Ctrl siRNA or siRNA targeting ATF4 mRNA. HSP90 was used as loading control. ( I ) mRNA expression of ASNS, CHAC1, CHOP, and SLC7A11 in F12-cultured A549 cells transfected with Ctrl siRNA or siRNA targeting ATF4 mRNA. GAPDH was used as a reference gene for normalization. ( J ) Western blotting and quantification of ATF4 and CHOP in protein extracts of F12-cultured A549 cells that were transfected with Ctrl siRNA or siRNA targeting ATF4 mRNA. HSP90 was used as loading control. ( K ) BSO dose response curves for A549 (left) and H838 (right) cells cultured in F12 medium for 72 h and transfected with Ctrl siRNA or siRNA targeting ATF4 mRNA. ( L ) Quantification by flow cytometry of oxidized BODIPY-C11 fluorescence in A549 (left) or H838 (right) cells that were cultured in F12 medium and treated with 100 μM BSO or vehicle for 24 h and transfected with Ctrl siRNA or siRNA targeting ATF4 mRNA. ( M ) Western blotting of CHOP in protein extracts of F12-cultured A549 (top) or H838 (bottom) cells that were transfected with Ctrl siRNA or siRNA targeting CHOP mRNA. HSP90 was used as loading control. ( N ) BSO dose response curves for A549 (left) and H838 (right) cells cultured in F12 medium for 72 h and transfected with Ctrl siRNA or siRNA targeting CHOP mRNA. ( O ) Western blotting of ATF4 and CHOP in protein extracts of F12-cultured A549 cells that carried a lentivirus overexpressing CHOP cDNA or control. HSP90 was used as loading control. ( P ) BSO dose response curves for A549 cells that carried lentivirus overexpressing CHOP cDNA or control and were cultured in F12 for 72 h. ( Q ) Western blotting of ATF4 and CHOP in protein extracts of RPMI-cultured A549 cells that carried a lentivirus overexpressing ATF4 cDNA or control. HSP90 was used as loading control. (R) BSO dose response curves for A549 cells that carried a lentivirus overexpressing ATF4 cDNA or control and were cultured in RPMI medium for 72 h. Dose response curves were normalized against the mean of the untreated samples for each condition. n = 3 replicates for all datapoints, error bars show SEM. ∗∗∗∗P < 0.0001, ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05.

Article Snippet: Cas9-expressing A549 cells were obtained from GeneCopoeia (SL-504; GeneCopoeia, Inc., Rockville, MD) and the mouse KP cell line was obtained from V. Sayin and is described [ ].

Techniques: Activation Assay, Western Blot, Cell Culture, Transfection, Control, Expressing, Flow Cytometry, Fluorescence

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: MagGO induces tumor cell death via frequency- and mode-dependent mechanical disruption (A) TEM images of MagGO synthesized under MF. The dashed white line represents the contour edge of GO. Scale bars, 200 nm. (B) M-H curve of GO, MNP, and MagGO. (C) AFM image of MagGO and the height of GO in MagGO. Scale bars, 500 nm. (D, F, and H) Cell viability of U87 (D), MDA-MB-231 (F), and A549 (H) cells treated with MNP and MagGO under RMF and 3D MF (RMF combined with OMF stimulation) at 5 Hz. The applied field strength is 75 mT. The data were presented as the mean ± SD. (E, G, and I) Cell viability of U87 (E), MDA-MB-231 (G), and A549 (I) cells treated with MNP and MagGO under 3D MF of different frequencies. The applied field strength is 75 mT. The data were presented as the mean ± SD.

Article Snippet: Glioblastoma U87 cells, human breast cancer cell line MDA-MB-231 cells, and human lung adenocarcinoma cell line A549 cells (American Type Culture Collection, Manassas, VA, USA) were cultured at 37°C under a humidified atmosphere containing 5% CO 2 with culture medium DMEM (Hyclone) containing of 1% penicillin and streptomycin (Hyclone) and 10% fetal bovine serum (FBS, Gibco).

Techniques: Disruption, Synthesized

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: MagGO induces lysosomal disruption (A) CLSM images of MNPs and MagGO in U87 cells. Lysosomes were stained with LysoTracker red (red), and the MNPs and MagGO were labeled with FITC (green). Scale bars, 15 μm. (B and C) Intensity profiles (white dashed line) of signals from lysosome and MNPs/MagGO fluorescent channels in (A). (D) CLSM images of MNPs and MagGO in MDA-MB-231 cells. Lysosomes were stained with LysoTracker red (red), and the MNPs and MagGO were labeled with FITC (green). Scale bars, 15 μm. (E and F) Intensity profiles (white dashed line) of signals from lysosomes and MNPs/MagGO fluorescent channels in (D). (G) CLSM images of MNPs and MagGO in A549 cells. Lysosomes were stained with LysoTracker red (red), and the MNPs and MagGO were labeled with FITC (green). Scale bars, 15 μm. (H and I) Intensity profiles (white dashed line) of signals from lysosomes and MNPs/MagGO fluorescent channels in (G). (J) CLSM images of U87 cells transfected with the EGFP-Gal3 plasmid after MagGO treatment under 3D MF. The applied field strength is 75 mT. The duration of magnetic field application is 30 min. Scale bars, 15 μm. (K) Counts of Gal3 puncta per U87 cell ( n = 10). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (L) Counts of Gal3 puncta per MDA-MB-231 cell ( n = 10). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (M) Counts of Gal3 puncta per A549 cell ( n = 10). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (N) Bio-TEM of lysosomal membrane morphology after mechanoporation for MagGO and MagGO+3D MF. The applied field strength is 75 mT. The duration of magnetic field application is 30 min. Scale bars, 1 μm. The dark blue arrow indicates the site of LMP, while the length of the blue arrow represents the size of the lysosomal membrane “wound”.

Article Snippet: Glioblastoma U87 cells, human breast cancer cell line MDA-MB-231 cells, and human lung adenocarcinoma cell line A549 cells (American Type Culture Collection, Manassas, VA, USA) were cultured at 37°C under a humidified atmosphere containing 5% CO 2 with culture medium DMEM (Hyclone) containing of 1% penicillin and streptomycin (Hyclone) and 10% fetal bovine serum (FBS, Gibco).

Techniques: Disruption, Staining, Labeling, Transfection, Plasmid Preparation, Membrane

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: MagGO primarily induces pyroptosis as the mode of cell death (A–C) CLSM images of CTSB release of U87 (A), MDA-MB-231 (B), and A549 (C) cells after MagGO treatment under 3D MF. The applied field strength is 75 mT. The duration of magnetic field application is 30 min. Scale bars, 20 μm. (D–F) The cell viabilities of U87 (D), MDA-MB-231 (E), and A549 (F) cells treated with MagGO were assessed after pre-treatment with z-VAD-FMK (10 μM), Necrostatin-1 (10 μM), 3-Methyladenine (10 μM), Ferrostatin-1 (2 μM), and MCC950 (10 nM) for 4 h, followed by exposure to 3D MF at 5 Hz for 30 min. The applied field strength was 75 mT. The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post hoc test. (G and H) Quantification of IL-1β (G) and IL-18 (H) release from U87 cells for control, MagGO, and MagGO+3D MF ( n = 3). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (I and J) Western blot analysis of Casp-1 (I) and GSDMD (J) in U87 cells for control, MagGO, and MagGO+3D MF.

Article Snippet: Glioblastoma U87 cells, human breast cancer cell line MDA-MB-231 cells, and human lung adenocarcinoma cell line A549 cells (American Type Culture Collection, Manassas, VA, USA) were cultured at 37°C under a humidified atmosphere containing 5% CO 2 with culture medium DMEM (Hyclone) containing of 1% penicillin and streptomycin (Hyclone) and 10% fetal bovine serum (FBS, Gibco).

Techniques: Control, Western Blot

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: Broad antitumor activity of MagGO across multiple tumor models (A) Schematic illustrations of in vivo anticancer therapy for MDA-MB-231 and A549 tumors. The applied field strength is 75 mT. The applied field frequency is 5 Hz. The duration of magnetic field application is 30 min. (B) The MDA-MB-231 tumor volume comparison of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF for 14 days ( n = 5). (C) The MDA-MB-231 tumor images of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). (D) The MDA-MB-231 tumor weight of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post hoc test. (E) The MDA-MB-231 tumor volume of each mouse in the control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF groups for 14 days. (F) The A549 tumor volume comparison of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF for 14 days ( n = 5). (G) The A549 tumor images of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). (H) The A549 tumor weight of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post hoc test. (I) The A549 tumor volume of each mouse in the control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF groups for 14 days.

Article Snippet: Glioblastoma U87 cells, human breast cancer cell line MDA-MB-231 cells, and human lung adenocarcinoma cell line A549 cells (American Type Culture Collection, Manassas, VA, USA) were cultured at 37°C under a humidified atmosphere containing 5% CO 2 with culture medium DMEM (Hyclone) containing of 1% penicillin and streptomycin (Hyclone) and 10% fetal bovine serum (FBS, Gibco).

Techniques: Activity Assay, In Vivo, Comparison, Control

Journal: Journal of Advanced Research

Article Title: Squalene epoxidase promotes paraquat-induced pulmonary toxicity through endoplasmic reticulum-mediated ferroptosis

doi: 10.1016/j.jare.2025.05.064

Figure Lengend Snippet: PQ induces ferroptosis in lung epithelial cells. (A) A549 cells were treated with PQ (0–2000 μM) and cell viability was assessed. (B-C) A549 cells treated with PQ (1000 μM, 0–24 h) (B) or PQ (0–1000 μM,24 h) (C) and the lipid ROS were measured using C11-BODIPY staining. (D-E) The labile iron pool was evaluated via Calcein-AM staining in A549 cells treated with PQ (1000 μM) (D) for 24 h (E). Images were captured with InCucyte. Bar = 400 µm. (F) MDA content in A549 cells. (G) Iron content in A549 cells. (H) Mito-tracker staining was evaluated for mitochondrial morphological change in A549 cells following 24-hour PQ (1000 μM) treatment. Bar = 10 µm. (I) Fe 2+ levels were measured by FerroOrange staining in A549 cells after 24-hour PQ (1000 μM) treatment. Bar = 10 µm. (J, K, L) Cells were treated with PQ (1000 μM) for 24 h in the presence or absence of DFO (50 μM), DFP (150 μM), RSL3 (1 μM), FeCl 3 (10 μM), FeSO 4 (10 μM) or FAC (10 μM) pretreatment for 1 h, and the cell viability was determined by CCK-8 assay. (M) Effect of PQ on GPX4 and SLC7A11 expression. Data are presented as Mean ± S.D. of at least three independent biological replicates (n = 3). *P < 0.05 , ** P < 0.01 , *** P < 0.001, **** P < 0.0001 show significant differences from each group .

Article Snippet: A549 human alveolar epithelial cells, RLE-6TN rat alveolar epithelial cells, and HEK-293 T cells were sourced from the American Type Culture Collection (ATCC, Manassas, VA, USA), while BEAS-2B human bronchial epithelial cells were obtained from Beyotime.

Techniques: Staining, CCK-8 Assay, Expressing

Journal: Journal of Advanced Research

Article Title: Squalene epoxidase promotes paraquat-induced pulmonary toxicity through endoplasmic reticulum-mediated ferroptosis

doi: 10.1016/j.jare.2025.05.064

Figure Lengend Snippet: ER stress mediates PQ-induced ferroptosis. (A-B) A549 cells were transfected with siRNA of PERK or NC (negative control) and then treated with PQ for 24 h. The protein expression (A) and the cell viability were determined (B). (C) Western blotting analysis of p-PERK, PERK, p-eIF2α, eIF2α, and CHOP in A549 cells pre-treated with 1 μM GSK157 for 1 h followed by PQ (1000 μM) treatment for 24 h. (D-E). A549 cells pre-treated with GSK157 (0.25, 0.5, 1 μM) for 1 h followed by PQ (1000 μM) treatment for 24 h, then ATP release (D) and CCK-8 assay (E) were determined. (F) Effects of GSK157 on PQ-induced lipid peroxidation. A549 Cells were pretreated with GSK157 for 1 h and then exposed to PQ for 24 h, followed by C11-BODIPY staining and determined by flow cytometry analysis. (G) Effects of GSK157 on PQ-induced iron content elevation. Cells were pretreated with GSK157 for 1 h and then exposed to PQ for 24 h, followed by calcein-AM staining and determined by flow cytometry analysis. (H) The staining of ER-Tracker (green) and FerroOrange (red) with Hoechst (blue) was photographed by the confocal microscope (Bar = 10 μm). (I) Iron content in A549 cells. (J) Effect of RSL3 on PQ-induced ER-stress. Cells were treated with PQ (1000 μM) for 24 h in the presence or absence of RSL3 (1 μM) and GSK157 (1 μM) pretreatment for 1 h. CCK-8 assay. (K) The expression levels of GPX4 and SLC7A11 were determined by Western blotting. Data are presented as Mean ± S.D. of at least three independent biological replicates (n = 3). *P < 0.05 , ** P < 0.01 , *** P < 0.001 show significant differences from each group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: A549 human alveolar epithelial cells, RLE-6TN rat alveolar epithelial cells, and HEK-293 T cells were sourced from the American Type Culture Collection (ATCC, Manassas, VA, USA), while BEAS-2B human bronchial epithelial cells were obtained from Beyotime.

Techniques: Transfection, Negative Control, Expressing, Western Blot, CCK-8 Assay, Staining, Flow Cytometry, Microscopy

Journal: Journal of Advanced Research

Article Title: Squalene epoxidase promotes paraquat-induced pulmonary toxicity through endoplasmic reticulum-mediated ferroptosis

doi: 10.1016/j.jare.2025.05.064

Figure Lengend Snippet: SQLE mediates PQ-induced lung epithelial ferroptosis. (A) Cells were transfected with siRNA of SQLE or NC and then treated with PQ for 24 h. Cell viability was evaluated by CCK-8 assay. (B) SQLE knockout cells on PQ-induced cell death were measured by CCK-8 assay. (C) The effect of PQ on A549 cells overexpressing SQLE (SQLE-WT) or SQLE mutant (SQLE-Y195F) cells was determined by CCK-8. The expression levels of SQLE in A549 (D) and BEAS-2B (E) were determined by Western blotting. (F-H) A549 SQLE-WT cells and A549 SQLE-Y195F cells were transfected with siRNA of SQLE or NC and then treated with PQ for 24 h was evaluated by CCK-8 assay (F), morphological changes. Bar = 400 µm. (G), and PI staining (H). Data are presented as Mean ± S.D. of at least three independent biological replicates (n = 3). *P < 0.05 , ** P < 0.01 , *** P < 0.001 show significant differences from each group. ns , no significance.

Article Snippet: A549 human alveolar epithelial cells, RLE-6TN rat alveolar epithelial cells, and HEK-293 T cells were sourced from the American Type Culture Collection (ATCC, Manassas, VA, USA), while BEAS-2B human bronchial epithelial cells were obtained from Beyotime.

Techniques: Transfection, CCK-8 Assay, Knock-Out, Mutagenesis, Expressing, Western Blot, Staining

Journal: Journal of Advanced Research

Article Title: Squalene epoxidase promotes paraquat-induced pulmonary toxicity through endoplasmic reticulum-mediated ferroptosis

doi: 10.1016/j.jare.2025.05.064

Figure Lengend Snippet: SQLE inhibition attenuates PQ-induced lung epithelial cell death by inhibiting ferroptosis. (A-C) A549 sgSQLE cells were treated with PQ (1000 μM). Confocal microscopy images show FerroOrange (red) and Hoechst (blue) staining (Bar = 10 μm) (A), GSH content was assessed (B), and Western blotting analysis evaluated GPX4 expression (C). (D) Cells were transfected with siRNA of SQLE or NC and then treated with PQ for 24 h. Western blotting analysis evaluates the expression of GPX4. (E-F) A549 sgSQLE cells were treated with PQ (1000 μM) for 24 h with or without pretreatment of RSL3 (1 μM) and FeCL3 (10 μM) for 1 h, measured by CCK-8. (G-I) A549-SQLE-WT and A549-SQLE-Y195F cells were treated with PQ (1000 μM) for 24 h with or without pretreatment of inhibitors (RSL3 1 μM, FeCL 3 10 μM, FeSO 4 10 μM) for 1 h, assessed by CCK-8. (J) HEK-293 T cells were transfected with the plasmid of SQLE-WT or SQLE-Y195F and then treated with PQ (1000 μM) for 24 h. Western blotting analysis evaluates the expression of SQLE and GPX4. Data are presented as Mean ± S.D. of at least three independent biological replicates (n = 3). *P < 0.05 , ** P < 0.01 , *** P < 0.001 show significant differences from each group. ns , no significance. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: A549 human alveolar epithelial cells, RLE-6TN rat alveolar epithelial cells, and HEK-293 T cells were sourced from the American Type Culture Collection (ATCC, Manassas, VA, USA), while BEAS-2B human bronchial epithelial cells were obtained from Beyotime.

Techniques: Inhibition, Confocal Microscopy, Staining, Western Blot, Expressing, Transfection, CCK-8 Assay, Plasmid Preparation

Journal: Journal of Advanced Research

Article Title: Squalene epoxidase promotes paraquat-induced pulmonary toxicity through endoplasmic reticulum-mediated ferroptosis

doi: 10.1016/j.jare.2025.05.064

Figure Lengend Snippet: SQLE inhibitors attenuate PQ-induced lung epithelial cell death. (A-G) Cells were treated with PQ for 24 h in the presence or absence of LNT postreatment for 30 mins. CCK-8 assay (A), ATP release (B) in A549 cells. CCK-8 assay (C), ATP release (D) in BEAS-2B cells. CCK-8 assay (E), ATP release (F) in RLE-6TN cells. CCK-8 assay (G) in AECⅡ. (H) Cells were treated with PQ for 24 h in the presence or absence of NB598 postreatment for 30 mins in A549 cells. Followed by CCK-8 assay. Morphological changes. Bar = 400 µm. (I), and PI staining (J). Data are presented as Mean ± S.D. of at least three independent biological replicates (n = 3). *P < 0.05 , ** P < 0.01 , *** P < 0.001 show significant differences from each group.

Article Snippet: A549 human alveolar epithelial cells, RLE-6TN rat alveolar epithelial cells, and HEK-293 T cells were sourced from the American Type Culture Collection (ATCC, Manassas, VA, USA), while BEAS-2B human bronchial epithelial cells were obtained from Beyotime.

Techniques: CCK-8 Assay, Staining

Journal: Journal of Advanced Research

Article Title: Squalene epoxidase promotes paraquat-induced pulmonary toxicity through endoplasmic reticulum-mediated ferroptosis

doi: 10.1016/j.jare.2025.05.064

Figure Lengend Snippet: LNT attenuates PQ-induced lung epithelial cell death via inhibiting ferroptosis. Cells were treated with PQ (1000 μM) for 30 mins and then post-treated with LNT for 24 h. followed by measurement C11-BODIPY in flow cytometry (A). Calcein-AM staining by captured with InCucyte. Bar = 400 µm (B) and measured by flow cytometry (C). (D) MDA content. (E) Mito tracker (green) with Hoechst (blue) was photographed by the confocal microscope. Bar = 10 μM. (F) FerroOrange (red) with Hoechst (blue) was photographed by the confocal microscope. Bar = 10 μM. (G) Iron content. (H-I) Cells were treated with PQ (1000 μM) for 30 mins in the presence or absence of inhibitors (RSL3, FeCL 3 , FeSO 4 , and FAC) for 1 h pretreatment, and then LNT (20 μM) posttreatment for 24 h in A549 cells. Cell viability was determined by CCK-8 assay. (J) Western blotting analysis of GPX4 and SLC7A11. Data are presented as Mean ± S.D. of at least three independent biological replicates (n = 3). *P < 0.05 , ** P < 0.01 , *** P < 0.001 show significant differences from each group. ns , no significance. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: A549 human alveolar epithelial cells, RLE-6TN rat alveolar epithelial cells, and HEK-293 T cells were sourced from the American Type Culture Collection (ATCC, Manassas, VA, USA), while BEAS-2B human bronchial epithelial cells were obtained from Beyotime.

Techniques: Flow Cytometry, Staining, Microscopy, CCK-8 Assay, Western Blot

Journal: Journal of Advanced Research

Article Title: Squalene epoxidase promotes paraquat-induced pulmonary toxicity through endoplasmic reticulum-mediated ferroptosis

doi: 10.1016/j.jare.2025.05.064

Figure Lengend Snippet: SQLE promotes PQ-induced lung injury via ferroptosis mediated by endoplasmic reticulum stress. (A) A549 sgSQLE cells were treated with PQ (1000 μM). ER-Tracker (green) and FerroOrange (red) with Hoechst (blue) were photographed by the confocal microscope (Bar = 10 μm). A549 Cells were treated with PQ (1000 μM) for 30 mins and then post-treated with LNT for 24 h. (B) The staining of ER-Tracker (green) and FerroOrange (red) with Hoechst (blue) was photographed by the confocal microscope (Bar = 10 μm). (C) The expression levels of p-PERK, PERK, p-eIF2α, and eIF2α were determined by Western blotting. (D) The expression levels of p-PERK, PERK, p-eIF2α, eIF2α, ATF4, and CHOP were determined by Western blotting. (E) Western blotting analysis for p-PERK, PERK, p-eIF2α, and ATF4 in lung tissue. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: A549 human alveolar epithelial cells, RLE-6TN rat alveolar epithelial cells, and HEK-293 T cells were sourced from the American Type Culture Collection (ATCC, Manassas, VA, USA), while BEAS-2B human bronchial epithelial cells were obtained from Beyotime.

Techniques: Microscopy, Staining, Expressing, Western Blot