Journal: iScience

Article Title: FBP17-mediated finger-like membrane protrusions in cell competition between normal and RasV12-transformed cells

doi: 10.1016/j.isci.2021.102994

Figure Lengend Snippet:

Article Snippet: The selective Cdc42 inhibitor ML141 (20 μM) and the antioxidant Trolox (1 mM) were from Tocris Bioscience and Cayman Chemical, respectively.

Techniques: Recombinant, Electron Microscopy, Software

Journal: Nature Communications

Article Title: Senescence-associated lysosomal dysfunction impairs cystine deprivation-induced lipid peroxidation and ferroptosis

doi: 10.1038/s41467-025-61894-9

Figure Lengend Snippet: a Presenescent TIG-3 cells (Control) were rendered senescent by 15 Gy X-ray irradiation (Irrad) or passaging (replicative senescence: Rep). These cells were then subjected to a viability assay after 24 h of culture within varying concentrations of erastin. b Measurement of intracellular free ferrous iron. Representative data from three independent experiments are shown. Scale bar, 100 µm. The box plot indicates the total intensity of FerroOrange staining. c – e Cell viability, LDH release and cytosolic ROS levels of control and senescent (Irrad and Rep) cells after 24 h ( c, d ) or 7 h ( e ) of treatment with 5 µM erastin alone or in combination with 500 nM ferrostatin-1 (Fer-1), 100 µM deferoxamine (DFO), 100 µM Trolox or 1 mM N-acetyl cysteine (NAC). f Quantification of the intracellular GSH content after 7 h of treatment with 5 µM erastin. g Lipid peroxidation assessed by C11-BODIPY fluorescence in control and senescent TIG-3 (Irrad and Rep) cells after 7 h of treatment with 5 µM erastin. The ratio of oxidized to reduced BODIPY (BODIPYox/BODIPYred) calculated from the median fluorescence intensity is shown. The data are presented as the mean ± s.d. of n = 3 biological replicates ( a , c , d , f , g ). For b , e the box represents the interquartile range (IQR), with a horizontal line and a black point indicating the median and mean, respectively. Whiskers extend to the most extreme data points within 1.5 times the IQR from the quartiles. White points beyond this range represent outliers. Statistical analysis was performed via two-way ANOVA with Tukey’s test ( a ), one-way-ANOVA with Tukey’s test ( c , d , f , g ), or the Kruskal-Wallis test followed by Dunn’s test ( b , e ). Source data are provided as a file.

Article Snippet: To induce ferroptosis, cells were cultured in medium containing erastin (Selleck, S7242) for 24 h. Ferroptosis inhibitors such as ferrostatin 1 (Fer-1; Selleck, S7243), Trolox (Selleck, S3665), DFO (Selleck, S5742), or NAC (Sigma, A9165) were coadministered to erastin-treated cells, followed by culture for 24 h. Concanamycin C (Adipogen Life Sciences, BVT-0254-C100) or EN6 (Selleck, S6650), which are compounds that alter lysosomal pH, were also coadministered with erastin, and the cells were incubated for 24 h.

Techniques: Control, Irradiation, Passaging, Viability Assay, Staining, Fluorescence

Journal: Nature Communications

Article Title: Senescence-associated lysosomal dysfunction impairs cystine deprivation-induced lipid peroxidation and ferroptosis

doi: 10.1038/s41467-025-61894-9

Figure Lengend Snippet: a Lysosomal pH of senescent TIG-3 (Irrad and Rep) cells after 24 h of treatment with or without 50 µM EN6, as measured by LysoSensor. b Measurement of free lysosomal ferrous iron (LysoRhoNox) and intracellular ferrous iron (FerroOrange) in senescent TIG-3 (Irrad and Rep) with or without 50 µM EN6 treatment. The box plot indicates the total intensity of LysoRhoNox staining. c Fluorescence images of lysosomes [LysoTracker (magenta)], lipid radicals [LipiRADICAL Green (green)], and nuclei [Hoechst 33342 (blue)] in senescent TIG-3 (Irrad and Rep) cells after treatment with 50 µM EN6. R represents the Pearson correlation coefficient between the signals of LysoTracker and LipiRADICAL Green. Scale bar, 50 µm. d Lipid peroxidation assessed by C11-BODIPY fluorescence in senescent TIG-3 cells (Irrad and Rep) with or without 2 µM erastin (Irrad), or 5 µM erastin (Rep) in the absence or presence of 50 µM EN6 for 12 h. The ratio of oxidized to reduced BODIPY (BODIPYox/BODIPYred) calculated from the median fluorescence intensity is shown. e Heatmap of peroxidized PC in senescent TIG-3 cells (Irrad) after 14 h of treatment with or without 5 µM erastin in the absence or presence of 50 µM EN6. f , g Cell viability of and LDH release by senescent TIG-3 (Irrad and Rep) cells after 24 h of treatment with or without 5 µM erastin in the absence or presence of 50 µM EN6 and with or without Fer-1 (500 nM), DFO (100 µM), Trolox (100 µM) or NAC (1 mM). h , i Senescent TIG-3 (Irrad and Rep) cells were infected with lentivirus encoding flag-tagged ATP6V1C2 or empty vector. After selection with puromycin, the cells were subjected to immunoblot analysis ( h ) or lysosomal pH analysis ( i ). j Viability of vector control or ATP6V1C2 overexpressing senescent TIG-3 cells (Irrad and Rep) after 24 h of treatment with 5 µM erastin alone or in combination with Fer-1 (500 nM), DFO (100 µM), Trolox (100 µM) or NAC (1 mM). k Mechanism of ferroptosis resistance in senescent cells. The data are presented as the mean ± s.d. of n = 3 biological replicates ( d , f , g , j ). For ( a , b , i ) the box represents the interquartile range (IQR), with a horizontal line and a black point indicating the median and mean, respectively. Whiskers extend to the most extreme data points within 1.5 times the IQR from the quartiles. White points beyond this range represent outliers. Statistical analysis was performed using the Kruskal-Wallis test followed by Dunn’s test ( a , b , i ) and one-way ANOVA with Tukey’s test ( d , f , g , j ). Source data are provided as a file.

Article Snippet: To induce ferroptosis, cells were cultured in medium containing erastin (Selleck, S7242) for 24 h. Ferroptosis inhibitors such as ferrostatin 1 (Fer-1; Selleck, S7243), Trolox (Selleck, S3665), DFO (Selleck, S5742), or NAC (Sigma, A9165) were coadministered to erastin-treated cells, followed by culture for 24 h. Concanamycin C (Adipogen Life Sciences, BVT-0254-C100) or EN6 (Selleck, S6650), which are compounds that alter lysosomal pH, were also coadministered with erastin, and the cells were incubated for 24 h.

Techniques: Staining, Fluorescence, Infection, Plasmid Preparation, Selection, Western Blot, Control

Journal: Nature Communications

Article Title: Senescence-associated lysosomal dysfunction impairs cystine deprivation-induced lipid peroxidation and ferroptosis

doi: 10.1038/s41467-025-61894-9

Figure Lengend Snippet: a – c PANC-1 cells were treated with control vehicle or 50 nM gemcitabine (GEM) for 10 days, after which cellular senescence was induced. a Lysosomal pH of presenescent and senescent PANC-1 cells after 24 h of treatment with or without 50 µM EN6. b , c Cell viability of and LDH release by PANC-1 and senescent PANC-1 cells after 24 h of treatment with or without 5 µM erastin in the absence or presence of 50 µM EN6 and with Fer-1 (500 nM), DFO (100 µM), Trolox (100 µM) or NAC (1 mM). d Schematic representation of the PANC-1 xenograft mouse model. e Relative tumor growth rates of PANC-1 xenografts after 34 days of treatment with vehicle or EN6 (50 mg/kg, i.p., every 3 days). The arrow indicates the date when the treatment was started. f Images of xenograft tumors harvested from mice. Scale bar, 1 cm. Tumor weight measurements of PANC-1 xenografts after 34 days of treatment with vehicle or EN6 (50 mg/kg, i.p., every 3 days). g Representative images of IHC staining of PANC-1 xenografts after 34 days of treatment with vehicle or EN6 (50 mg/kg, i.p., every three days). Scale bar, 100 µm. The plot shows the percentage of the positive area of 4-HNE and Ptsg2 IHC staining in vehicle- and EN6-treated PANC-1 tumors. Data are represented as mean ± s.d For ( b , c ) the data are presented as the mean ± s.d. of n = 3 biological replicates. For a the box represents the interquartile range (IQR), with a horizontal line and a black point indicating the median and mean, respectively. Whiskers extend to the most extreme data points within 1.5 times the IQR from the quartiles. White points beyond this range represent outliers. For ( e , f ) the data are presented as the mean ± s.d. of n = 6 (Vehicle), n = 7 (EN6) biological replicates. Statistical analysis was performed using the Kruskal-Wallis test followed by Dunn’s test ( a ), one-way-ANOVA with Tukey’s test ( b , c ), and the two-sided Mann-Whitney U test ( e , f , g ). Source data are provided as a file.

Article Snippet: To induce ferroptosis, cells were cultured in medium containing erastin (Selleck, S7242) for 24 h. Ferroptosis inhibitors such as ferrostatin 1 (Fer-1; Selleck, S7243), Trolox (Selleck, S3665), DFO (Selleck, S5742), or NAC (Sigma, A9165) were coadministered to erastin-treated cells, followed by culture for 24 h. Concanamycin C (Adipogen Life Sciences, BVT-0254-C100) or EN6 (Selleck, S6650), which are compounds that alter lysosomal pH, were also coadministered with erastin, and the cells were incubated for 24 h.

Techniques: Control, Immunohistochemistry, MANN-WHITNEY

Journal: Viruses

Article Title: In-Gel Isolation and Characterization of Large (and Other) Phages

doi: 10.3390/v12040410

Figure Lengend Snippet: Plots of plaque diameter vs. A for phages (adapted from of reference ). Key: 36, phage 0305phi8-36; T4, phage T4; G, Phage G.

Article Snippet: However, when comparing jumbo myophage G to jumbo myophage 0305phi8-36 (218.948 Kb genome [ ]; [ ]), this relationship is lost in that one visually observes no significant difference in slope, even though (1) the surface area of phage G (cryo-EM data in [ ]) is over 2x the surface area of phage 0305phi8-36 (cryo-EM data in [ ]) and (2) gel sieving is best correlated with particle surface area [ ].

Techniques: