jasplakinolide  (Tocris)

Journal: Advanced Science

Article Title: ERM Inhibition Confers Ferroptosis Resistance through ROS‐Induced NRF2 Signaling

doi: 10.1002/advs.202513310

Figure Lengend Snippet: ERM‐Actin axis regulates erastin‐induced ferroptosis. (a) Representative confocal microscopy images of phalloidin‐stained HT‐1080 cells treated with DMSO or erastin (5 µ m ) for 10 h. Images were acquired as SUM projections using a confocal microscopy with a 60x objective, capturing data through the FITC channel. The image on the right shows an enlarged view of the area within the red box. Confocal imaging was repeated twice ( n > 30). (b) Quantification of fluorescence intensity in HT‐1080 cells from a ( n = 46). (c) Representative confocal microscopy images of HT‐1080 cells stained with phalloidin (F‐actin) and anti‐Ezrin antibody. Upper graphs display straightened lines along the cell periphery (white dashed arrow), and lower panels show magnified views of regions marked by white rectangles. Images were acquired as MAX projections using a 60x objective. Confocal imaging was repeated twice ( n > 30). (d) Time‐lapse imaging of LifeAct‐mScarletI‐expressed HT‐1080 cells treated with DMSO, NSC305787 (2 µ m ), or NSC668394 (5 µ m ). Confocal imaging was performed twice ( n > 30). The time point marked as 0 min represents the start of the movie, with a ∼15–30 min gap between chemical treatment and the movie start. (e) Cell death measurement of HT‐1080 cells treated with LatA (0.05 and 0.1 µg/mL) and Erastin (5 µ m ) for 30 h. Dead cells were labeled with Propidium iodide. (f) Cell death measurement of HeLa cells treated with CytoD (0.125, 0.5, and 1 µ m ) and Erastin (10 µ m ) for 14 h. Dead cells were labeled with Propidium iodide. (g) Cell death measurement of HT‐1080 cells treated with CytoD (0.25, 0.5, 1, and 2 µ m ) and Erastin (5 µ m ) for 20 h. Dead cells were labeled with Propidium iodide. (h) Cell death measurement of HT‐1080 cells treated with the indicated combination of Erastin (5 µ m ), NSC305787 (2 µ m ), and LatA (0.05 and 0.1 µg/mL) for 23 h. Dead cells were labeled with Propidium iodide. (i) Cell death measurement of HT‐1080 cells treated with the indicated combination of Erastin (5 µ m ), NSC668394 (5 µ m ), and LatA (0.05 and 0.1 µg/mL) for 23 h. Dead cells were labeled with Propidium iodide. (j) Cell death measurement of shERM HT‐1080 cells treated with the indicated combination of Erastin (5 µ m ) and LatA (0.05 µg/mL) for 18 h. Dead cells were labeled with Propidium iodide. (k) Cell death measurement of HT‐1080 cells treated with the indicated combination of Erastin (5 µ m ), NSC305787 (2 µ m ), and jasplakinolide (40 n m ) for 18 h. Dead cells were labeled with Propidium iodide. (l) Cell death measurement of HT‐1080 cells treated with the indicated combination of Erastin (5 µ m ), NSC668394 (5 µ m ), and jasplakinolide (40 n m ) for 36 h. Dead cells were labeled with Propidium iodide. (m) Cell death measurement of shERM HT‐1080 cells treated with the indicated combination of Erastin (5 µ m ) and jasplakinolide (40 n m ) for 22 h. Dead cells were labeled with Propidium iodide. Data and error bars are mean ± SEM, n = 3 biologically independent experiments in e–m. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; n.s., not significant. All p values were calculated using a one‐way or two‐way analysis of variance (ANOVA).

Article Snippet: NSC305787 (MCE, #HY‐18931A) (dissolved in DMSO, sonicated for 5 min to make a 10 m m stock solution, aliquoted and stored at −80°C, re‐sonicated for 2 min before use); NSC668394 (MCE, #HY‐115492); Erastin (Sellcek, #S7242) (dissolved in DMSO to make a 10 m m stock solution, aliquoted and stored at −80°C, avoid repeated freeze‐thaw cycles); Jasplakinolide (SANTN CRUZ, #102396‐24‐7); ML385 (TargetMol, #T4360); Zinc Protoporphyrin IX (ZnPP, MCE, #HY‐101193); (1S, 3R)‐RSL3 (Selleck, #S8155); ML210 (MCE, #HY‐100003); Cytochalasin D (CytoD, Glpbio, #GC13440); tert‐butyl hydroperoxide (tBOOH, MACKLIN, #B802372‐50 mL); 2,3‐dimethoxy‐1,4‐naphthalenedione (DMNQ, MCE, #HY‐121026); MG‐132 (MCE, #HY‐13259); Z‐VAD (MCE, #HY‐16658B); CuCl 2 (Macklin, #C804816); Elesclomol (Macklin, #E864529); Ammonium tetrathiomolybdate (TTM, Macklin, #A828261); SB‐663825 (MCE, HY‐108333); SLK/STK10‐IN‐1 (MCE, #HY‐132868); Chloroquine (CQ, MCE, #HY‐17589A); CK‐666 (MCE, #HY‐16926); Ferrostatin‐1 (Fer‐1, MCE, #HY‐100579); Latrunculin A (LatA, Abcam, #ab144290); Brusatol (MCE, #HY‐19543); Liproxstatin‐1 (Lip‐1, Macklin, #950455‐15‐9); Deferoxamine mesylate (DFO, MCE, #HY‐B0988); 3% H 2 O 2 (Lircon, #6926378903443); N‐acetylcysteine (NAC, Sigma, #A7250); Cisplatin (CDDP, Selleck, #S1166).

Techniques: Confocal Microscopy, Staining, Imaging, Fluorescence, Labeling

Journal: Advanced Science

Article Title: ERM Inhibition Confers Ferroptosis Resistance through ROS‐Induced NRF2 Signaling

doi: 10.1002/advs.202513310

Figure Lengend Snippet: ERM inhibition induces actin‐dependent ROS elevation. (a) ROS levels measured by DCFH‐DA staining in HT‐1080 cells treated with DMSO, NSC305787 (2 µ m ) over the indicated time course. (b) Quantification of ROS levels from a. (c) ROS levels measured by DCFH‐DA staining in HT‐1080 cells treated with DMSO, NSC668394 (5 µ m ) over the indicated time course. (d) Quantification of ROS levels from c. (e) ROS levels measured by DCFH‐DA staining in HT‐1080 subjected to individual or combined treatments with NSC305787 (2 µ m ), NSC668394 (5 µ m ), or NAC (500 µ m ) for 3 h. (f) Quantification of ROS levels from e. (g) ROS levels measured by DCFH‐DA staining in HT‐1080 cells treated with DMSO, Erastin (5 µ m ) over the indicated time course. (h) Quantification of ROS levels from g. (i) ROS levels measured by DCFH‐DA staining in HT‐1080 cells pre‐treated with LatA (0.1 µg/mL) for 2 h following by co‐treatment with LatA (0.1 µg/mL) and either NSC305787 (2 µ m ) or NSC668394 (5 µ m ) for an additional 2 h. (j) Quantification of ROS levels from i. (k) ROS levels measured by DCFH‐DA staining in shEzrin‐2, shRadixin‐1, shMoesin‐2 HT‐1080 cells treated with LatA (0.05 µg/mL) for 3 h. (l) Quantification of ROS levels from k. (m) ROS levels measured by DCFH‐DA staining in HT‐1080 cells pre‐treated with jasplakinolide (50 n m ) for 2 h followed by co‐treatment with jasplakinolide (50 n m ) and either NSC305787 (2 µ m ) or NSC668394 (5 µ m ) for an additional 2 h. (n) Quantification of ROS levels from m. (o) ROS levels measured by DCFH‐DA staining in shEzrin‐2, shRadixin‐1, shMoesin‐2 HT‐1080 cells treated with jasplakinolide (50 n m ) for 4 h. (p) Quantification of ROS levels from o. (q) Superoxide anion levels measured by dihydroethidium (DHE) staining in HT‐1080 cells treated with DMSO, NSC305787 (2 µ m ), or NSC668394 (5 µ m ) for 4 h. (r) Quantification of ROS levels from q. (s) Schematic showing AlphaFold3‐predicted structures of Ezrin, actin, and NOX2. (t) ROS levels measured by DCFH‐DA staining in HT‐1080 cells pre‐treated with Apocynin (20 µ m ) for 2 h followed by co‐treatment with Apocynin (20 µ m ) and either NSC305787 (2 µ m ) or NSC668394 (5 µ m ) for an additional 2 h. (u) Quantification of ROS levels from t. (v) ROS levels measured by DCFH‐DA staining in shEzrin‐2, shRadixin‐1, shMoesin‐2 HT‐1080 cells treated with Apocynin (20 µ m ) for 4 h. (w) Quantification of ROS levels from v. Data and error bars are mean ± SEM, n = 3 biologically independent experiments in b, d, f, h, j, l, n, p, r, u, and w. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; n.s., not significant. All p values were calculated using a one‐way or two‐way analysis of variance (ANOVA).

Article Snippet: NSC305787 (MCE, #HY‐18931A) (dissolved in DMSO, sonicated for 5 min to make a 10 m m stock solution, aliquoted and stored at −80°C, re‐sonicated for 2 min before use); NSC668394 (MCE, #HY‐115492); Erastin (Sellcek, #S7242) (dissolved in DMSO to make a 10 m m stock solution, aliquoted and stored at −80°C, avoid repeated freeze‐thaw cycles); Jasplakinolide (SANTN CRUZ, #102396‐24‐7); ML385 (TargetMol, #T4360); Zinc Protoporphyrin IX (ZnPP, MCE, #HY‐101193); (1S, 3R)‐RSL3 (Selleck, #S8155); ML210 (MCE, #HY‐100003); Cytochalasin D (CytoD, Glpbio, #GC13440); tert‐butyl hydroperoxide (tBOOH, MACKLIN, #B802372‐50 mL); 2,3‐dimethoxy‐1,4‐naphthalenedione (DMNQ, MCE, #HY‐121026); MG‐132 (MCE, #HY‐13259); Z‐VAD (MCE, #HY‐16658B); CuCl 2 (Macklin, #C804816); Elesclomol (Macklin, #E864529); Ammonium tetrathiomolybdate (TTM, Macklin, #A828261); SB‐663825 (MCE, HY‐108333); SLK/STK10‐IN‐1 (MCE, #HY‐132868); Chloroquine (CQ, MCE, #HY‐17589A); CK‐666 (MCE, #HY‐16926); Ferrostatin‐1 (Fer‐1, MCE, #HY‐100579); Latrunculin A (LatA, Abcam, #ab144290); Brusatol (MCE, #HY‐19543); Liproxstatin‐1 (Lip‐1, Macklin, #950455‐15‐9); Deferoxamine mesylate (DFO, MCE, #HY‐B0988); 3% H 2 O 2 (Lircon, #6926378903443); N‐acetylcysteine (NAC, Sigma, #A7250); Cisplatin (CDDP, Selleck, #S1166).

Techniques: Inhibition, Staining

Journal: bioRxiv

Article Title: Amplified MS2 labeling reveals heterogeneous dynamics of RNA granules in the live murine brain

doi: 10.64898/2026.02.03.703574

Figure Lengend Snippet: Movements of neuronal RNA granules. (a), (b) Single frame, maximum and average projections of time-lapse images of MCP-4×sfGFP expressing cells in the Actb MBS/MBS ;Grik4-Cre hippocampus (time=162 sec). Arrowheads, cytoplasmic compartments limiting the access of RNPs. (c) Max projections of time-lapse images before and after the treatments of Jasplakinolide (10 µM), nocodazole (33 µM), and tetracaine (5 mM). (d) Single particle tracking traces overlaid with average projection. (time=33.8 sec). (e) Jump distance distribution with the best fit to two-component 2D diffusion model (red). Dashed lines, each component. Scale bars, 10 µm.

Article Snippet: Jasplakinolide and nocodazole (Santa Cruz Biotechnology, sc-202191 and sc-3518B, respectively) were dissolved in DMSO and added to the ACSF.

Techniques: Expressing, Single-particle Tracking, Diffusion-based Assay