Journal: Cell Death Discovery

Article Title: Neddylation inhibition induces DNA double-strand breaks, hampering tumor growth in vivo, and promotes radiosensitivity in PAX3–FOXO1 rhabdomyosarcoma

doi: 10.1038/s41420-025-02787-0

Figure Lengend Snippet: A Representative western blot ( n = 3 independent experiments) of the indicated proteins on RH4 and RH30 cells treated for 24 h with either vehicle (DMSO) or MLN4924 GI 50 , and then irradiated or not with a single dose of 4 Gy. Cells were assayed 6 h post-irradiation. Vinculin and αTubulin are the loading controls. B Representative images of RH4 and RH30 colonies treated as in ( A ). Colonies were stained with crystal violet 12 days post-cell seeding. C Histograms depict the number of colonies of RH4 and RH30 cells treated as in ( A ), calculated as a fold increase over vehicle (DMSO). Graph represents the mean ± SD ( n = 3 independent experiments). One-way ANOVA. Exact p -values are reported in the figure. D RH4 and RH30 cells were treated for 24 h with either vehicle (DMSO) or MLN4924 GI 50 and then irradiated or not with a single dose of 4 Gy and assayed after 24 h. Histograms depict the percentage of cells in G0/G1, S, and G2/M phases. Graph represents the mean ± SEM ( n = 3 independent experiments). Two-way ANOVA. Exact p -values are reported in the figure. Black p -values: vs DMSO; green p -values: vs MLN4924; yellow p -values: vs 4 Gy. E Representative diagrams of cell cycle flow cytometry analysis on RH4 and RH30 cells treated as in ( D ). F Histograms depict the percentage of Caspase 3/7 activity of RH4 and RH30 cells treated as in ( D ), calculated as fold increase over vehicle (DMSO, 100%). Graph represents the mean ± SEM ( n = 3 independent experiments). One-way ANOVA. Exact p -values are reported in the figure. G Histograms depict the percentage of Annexin-V positive/PI single- and double-positive RH4 and RH30 cells treated as in ( D ). Graph represents the mean ± SEM ( n = 3 independent experiments). One-way ANOVA. Exact p -values are reported in the figure. H Representative plots of flow cytometry analysis of Annexin V/PI staining of RH4 and RH30 cells treated as in ( D ). I Representative western blot ( n = 3 independent experiments) of the indicated proteins on RH4 and RH30 cells treated as in ( D ). αTubulin and Vinculin are the loading controls.

Article Snippet: Antibodies against SKP2 (H-435, sc-7164, 1:500), DNA-PKcs (sc-390849), p27 (sc-1641) were obtained from Santa Cruz Biotechnology Inc., (Santa Cruz, CA, USA); Vinculin (hVIN-1, V9131, 1:5000) was purchased from Sigma-Aldrich (St. Louis, MO, USA); MYOG (F5D-c, 1:200) was from DSHB (University of Iowa, Iowa City, IA, USA); α-TUBULIN (NB100-680, 1:5000) was from Novus Biologicals (Littleton, CO, USA).

Techniques: Western Blot, Irradiation, Staining, Flow Cytometry, Activity Assay

Journal: Cell Death Discovery

Article Title: Neddylation inhibition induces DNA double-strand breaks, hampering tumor growth in vivo, and promotes radiosensitivity in PAX3–FOXO1 rhabdomyosarcoma

doi: 10.1038/s41420-025-02787-0

Figure Lengend Snippet: A Representative western blot ( n = 3 independent experiments) of the indicated proteins on RH4 and RH30 cells treated for 24 h with either vehicle (DMSO) or MLN4924 GI 50 and then irradiated or not with a single dose of 4 Gy and assayed 6 h post-irradiation. Vinculin and αTubulin are the loading controls. B Representative immunofluorescence images of γH2AX (green) on RH4 and RH30 cells treated as in ( A ). Dapi (blue) was the nuclear counterstain. Images were acquired with a fluorescence microscope equipped with a 60× oil immersion objective. Scale bar = 25 μm. C Scatter dot plots depict the intensity of γH2AX staining per nucleus on RH4 and RH30 cells treated as in ( A ). The graph represents the mean ± SEM. Kruskal-Wallis test. Exact p -values are reported in the figure. D Representative immunofluorescence images of the neutral Comet assay on RH4 and RH30 cells treated as in ( A ) and assayed 24 h post-irradiation. Images were acquired with a fluorescence microscope equipped with a 40× oil immersion objective. Scale bar = 50 μm. E Scatter dot plots represent the mean of the tail moment ± SEM. One-way ANOVA. Exact p -values are reported in the figure.

Article Snippet: Antibodies against SKP2 (H-435, sc-7164, 1:500), DNA-PKcs (sc-390849), p27 (sc-1641) were obtained from Santa Cruz Biotechnology Inc., (Santa Cruz, CA, USA); Vinculin (hVIN-1, V9131, 1:5000) was purchased from Sigma-Aldrich (St. Louis, MO, USA); MYOG (F5D-c, 1:200) was from DSHB (University of Iowa, Iowa City, IA, USA); α-TUBULIN (NB100-680, 1:5000) was from Novus Biologicals (Littleton, CO, USA).

Techniques: Western Blot, Irradiation, Immunofluorescence, Fluorescence, Microscopy, Staining, Neutral Comet Assay

Journal: PLOS Genetics

Article Title: Bendless is essential for PINK1-Park mediated Mitofusin degradation under mitochondrial stress caused by loss of LRPPRC

doi: 10.1371/journal.pgen.1010493

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: Rabbit Tubulin , Novus Biologicals- NB100-56459 , 1:1000 WB.

Techniques:

Journal: PLOS Genetics

Article Title: Bendless is essential for PINK1-Park mediated Mitofusin degradation under mitochondrial stress caused by loss of LRPPRC

doi: 10.1371/journal.pgen.1010493

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: Rabbit Tubulin , Novus Biologicals- NB100-56459 , 1:1000 WB.

Techniques:

Journal: PLOS Genetics

Article Title: Bendless is essential for PINK1-Park mediated Mitofusin degradation under mitochondrial stress caused by loss of LRPPRC

doi: 10.1371/journal.pgen.1010493

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: Rabbit Tubulin , Novus Biologicals- NB100-56459 , 1:1000 WB.

Techniques:

Journal: Cells

Article Title: RNA-Sequencing Reveals Upregulation and a Beneficial Role of Autophagy in Myoblast Differentiation and Fusion.

doi: 10.3390/cells11223549

Figure Lengend Snippet: Figure 4. Effects of chloroquine (CQ) on LC3-I and LC3-II protein expression in C2C12 cells during myogenic differentiation. C2C12 cells were cultured in differentiation medium in the presence (+) or absence (−) of 20 µM CQ. Total protein lysates from cells on days 0 (D0, before differentiation), 1 (D1), 2 (D2), and 4 (D4) of differentiation were analyzed by Western blotting, using antibodies specific to LC3 and β-Tubulin (as a loading control). (A) Representative images of Western blots. (B) Ratio of LC3-II to LC3-I protein. ** indicates p < 0.002 (n = 5), based on t-tests.

Article Snippet: The following primary antibodies were used in Western blot analyses: ß-tubulin at 1:1000 dilution (E7 from DSHB), LC3B at 1:1000 dilution (NB100-2220 from Novus Biologicals, Centennial, CO, USA), and myogenin at 1:25 dilution (F5D from DSHB).

Techniques: Expressing, Cell Culture, Western Blot, Control

Journal: Cell metabolism

Article Title: Dissociation of Adaptive Thermogenesis from Glucose Homeostasis in Microbiome-Deficient Mice

doi: 10.1016/j.cmet.2020.01.012

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Beta tubulin , Novus Biologicals , Cat# NB100-56459; IMG5810A, RRID: AB_1152679.

Techniques: Virus, Recombinant, Sequencing, Software, Mass Spectrometry, Irradiation