Journal: Molecular Biology of the Cell

Article Title: Image-based drug screen identifies HDAC inhibitors as novel Golgi disruptors synergizing with JQ1

doi: 10.1091/mbc.E17-03-0176

Figure Lengend Snippet: Image-based chemical compound screen identifies HDAC inhibitors and DNA-damaging agents as novel Golgi-dispersing compounds. To identify novel compounds that modulate Golgi morphology, a screening platform was established. (A) Screening pipeline including cell seeding (A549 cells), treatment with compound library, staining for the cis -Golgi (GM130), the nucleus (Hoechst), the ER stress marker (GRP78), cytoplasm (Phalloidin), and image acquisition and processing. (B) Representative images of the negative control (vehicle-treated cells) as well as positive controls (BFA, doxorubicin, and nocodazole) and newly discovered Golgi-fragmenting drugs (Bleomycin, Vorinostat, 4-iodo-SAHA, Trichostatin A, Givinostat, and Pracinostat) are displayed. (C) Following image analysis, to exclude potential plate effects, the Golgi area of cells treated with the chemical library was normalized to the vehicle-treated sample present within the same plate. The corresponding survival ratios of treated cells are also shown. (D) Detailed view of compounds are shown, of which at least two of three replicates caused a ≥1.5-fold increase in Golgi area. The compound panel includes positive controls and novel Golgi-fragmenting compounds, which were selected for further investigation. *** p < 0.001 vs. #; see Materials and Methods .

Article Snippet: Compounds were obtained from the following companies: brefeldin A (Sigma-Aldrich), golgicide A (Santa Cruz Biotechnology), monensin (Enzo Life Sciences), AG-1478 (Sigma), tunicamycin (Santa Cruz Biotechnology), thapsigargin (Santa Cruz Biotechnology), nocodazole (Santa Cruz Biotechnology), (+)-JQ1 (Cayman Chemical), CBP30 (TargetMol), doxorubicin (Sigma), etoposide (Sigma), teniposide (Santa Cruz Biotechnology), mitomycin-C (Santa Cruz Biotechnology), cisplatin (Santa Cruz Biotechnology), hydroxyurea (Sigma), 5-fluorouracil (Sigma), gemcitabine (Santa Cruz Biotechnology), irinotecan (Santa Cruz Biotechnology), bleomycin (Santa Cruz Biotechnology), NU7441 (Selleckchem), KU55933 (Sigma), Flavopiridol (Santa Cruz Biotechnology), phorbol 12-myristate 13-acetate (PMA; Santa Cruz Biotechnology), Panobinostat (Selleckchem), Tubastatin (Selleckchem), Entinostat (Santa Cruz Biotechnology), Pracinostat (Selleckchem), Givinostat (Selleckchem), Triptolide (Santa Cruz Biotechnology), α-Amanitin (Santa Cruz Biotechnology), and Z-VAD-FMK (Santa Cruz Biotechnology).

Techniques: Drug discovery, Staining, Marker, Negative Control

Journal: Stem Cell Research & Therapy

Article Title: A scalable platform for EPSC-Induced MSC extracellular vesicles with therapeutic potential

doi: 10.1186/s13287-025-04507-y

Figure Lengend Snippet: Therapeutic effects of iMSC-EVs and MSC-EVs in a bleomycin-induced pulmonary fibrosis mouse model ( A ) Schematic representation of the experimental design. Pulmonary fibrosis was induced in mice by intratracheal instillation of bleomycin (5 mg/kg) on Day 0. Mice were treated every two days from Day 7 to Day 21 with either iMSC-EVs (20 µg/100 µL), MSC-EVs (20 µg/100 µL), or DPBS (control) before being sacrificed for analysis. ( B ) Body weight changes over time. Mice treated with iMSC-EVs or MSC-EVs exhibited significantly improved weight recovery compared to the BLM + PBS group, indicating a reduction in disease severity. Data are presented as mean ± SD. ( C ) Hematoxylin and eosin ( H & E ) staining of lung tissue. Representative images show severe alveolar structure disruption and fibrosis in the BLM + PBS group, whereas both iMSC-EVs and MSC-EVs treatments preserved lung architecture and reduced fibrotic lesions. Scale bar: 2.5 mm. ( D ) Masson’s trichrome staining for collagen deposition. The BLM + PBS group exhibited extensive collagen accumulation (blue staining), whereas both EV-treated groups showed reduced collagen deposition, suggesting attenuation of fibrosis. Scale bar: 2.5 mm. ( F ) Quantification of fibrosis severity using the Ashcroft score. Both iMSC-EVs and U iMSC-EVs treatments significantly decreased fibrosis scores compared to the BLM + PBS group, with no significant difference between the two EV-treated groups, indicating comparable therapeutic efficacy. Data are presented as mean ± SD. ( G ) Total protein levels in bronchoalveolar lavage fluid (BALF). EV-treated mice exhibited significantly lower BALF protein levels compared to the BLM + PBS group, suggesting reduced alveolar-capillary barrier damage and inflammation. Data are presented as mean ± SD

Article Snippet: Under 1.5% isoflurane (Lunan Better Phamacertical, 110302) anesthesia, mice received a single intratracheal instillation of bleomycin sulfate (2 mg/kg; MedChemExpress, HY-17565).

Techniques: Control, Staining, Disruption, Drug discovery

Journal: Mutagenesis

Article Title: Functional analysis of the new barley gene HvKu80 indicates that it plays a key role in double-strand DNA break repair and telomere length regulation.

doi: 10.1093/mutage/gev033

Figure Lengend Snippet: Figure 4. Relative HvKu80 gene expression levels in the parent cultivar ‘Sebastian’ (A), mutant ku80.c (B) and mutant ku80.j (C) after the treatment with bleomycin (100 µg/ml) and in the untreated material were analysed at various time points. ‘C’: seedlings after 36 h of growth at 24°C, ‘0’ in the untreated material denotes 36-h-old seedlings of barley that were treated with sterile water for 2 h at 37°C, ‘0’ in the treated material denotes 36-hold seedlings of barley that were treated with bleomycin for 2 h at 37°C. The value of the relative expression level was normalised to the standard control (HvARF1 gene, accession no. AJ508228.2). The expression level (2−ΔΔCt) in the untreated material was calculated relative to the control roots (C). The expression level (2−ΔΔCt) in the treated material was calculated relative to the untreated material at a particular time point. #Statistically significant difference vs. the control (C) (P < 0.05); *Statistically significant difference compared to the untreated material at the corresponding time point (P < 0.05).

Article Snippet: After this time, the seeds were incubated at 24°C for 36 h. Thirty-six-hour-old barley seedlings were transferred to 100 μg/ ml solution of bleomycin A5 hydrochloride (LKT Laboratories) and incubated at 37°C for 2 h. Upon the mutagenic treatment the seedlings were washed three times in sterile water for 15 min.

Techniques: Gene Expression, Mutagenesis, Sterility, Expressing, Control

Journal: Mutagenesis

Article Title: Functional analysis of the new barley gene HvKu80 indicates that it plays a key role in double-strand DNA break repair and telomere length regulation.

doi: 10.1093/mutage/gev033

Figure Lengend Snippet: Figure 5. The number of DSBs that were induced by bleomycin. Ethidium bromide-stained agarose gel visualising the induction of DSBs by bleomycin treatment in barley genomic DNA (A), concentration-dependent increase in DSB number in roots of the cultivar ‘Sebastian’ (B), the number of DSBs that were induced by 100 µg/ml bleomycin in the roots of the parent cultivar ‘Sebastian’ and the mutants ku80 after the bleomycin treatment (C). The DSB frequency was calculated for whole roots of the analysed genotypes. ‘M’: marker size 1 kb, ‘C’: measurement of DSB number was done on DNA isolated from whole roots after 36 h of growth at 24°C, ‘0’-treated material: measurement of DSB number was done on DNA isolated from whole 36-h-old roots of barley that were treated with bleomycin for 2 h at 37°C, *Statistically significant difference in relation to the parent cultivar ‘Sebastian’ (P < 0.05) at corresponding time points, #Statistically significant difference in relation to 0 µg/ml concentration of bleomycin (P < 0.05).

Article Snippet: After this time, the seeds were incubated at 24°C for 36 h. Thirty-six-hour-old barley seedlings were transferred to 100 μg/ ml solution of bleomycin A5 hydrochloride (LKT Laboratories) and incubated at 37°C for 2 h. Upon the mutagenic treatment the seedlings were washed three times in sterile water for 15 min.

Techniques: Staining, Agarose Gel Electrophoresis, Concentration Assay, Marker, Isolation

Journal: Mutagenesis

Article Title: Functional analysis of the new barley gene HvKu80 indicates that it plays a key role in double-strand DNA break repair and telomere length regulation.

doi: 10.1093/mutage/gev033

Figure Lengend Snippet: Figure 6. Number of MCD nuclei after the treatment of the roots of the cultivar ‘Sebastian’ and the mutants ku80.c and ku80.j with bleomycin. ‘C’: seedlings after 36 h of growth at 24°C, ‘0’-treated material: 36-h-old barley seedlings that were treated with bleomycin for 2 h at 37°C; *Statistically significant difference in relation to the parent cultivar ‘Sebastian’ (P < 0.05) at corresponding time points.

Article Snippet: After this time, the seeds were incubated at 24°C for 36 h. Thirty-six-hour-old barley seedlings were transferred to 100 μg/ ml solution of bleomycin A5 hydrochloride (LKT Laboratories) and incubated at 37°C for 2 h. Upon the mutagenic treatment the seedlings were washed three times in sterile water for 15 min.

Techniques:

Journal: Mutagenesis

Article Title: Functional analysis of the new barley gene HvKu80 indicates that it plays a key role in double-strand DNA break repair and telomere length regulation.

doi: 10.1093/mutage/gev033

Figure Lengend Snippet: Figure 7. Reduced growth of seedlings from the cultivar ‘Sebastian’ and the mutants ku80.c and ku80.j after treatment with bleomycin (100 µg/ml). Representative image of seedlings from the cultivar ‘Sebastian’ and the mutants ku80.c and ku80.j after treatment with bleomycin (A). Reduction of roots and shoots length of cultivar ‘Sebastian’ and the mutants ku80.c and ku80.j after bleomycin treatment (B). Control; ‘C’: seedlings after 60 h of growth at 24°C; scale = 3 cm, *Statistically significant difference in relation to root or shoot in control conditions (P < 0.05).

Article Snippet: After this time, the seeds were incubated at 24°C for 36 h. Thirty-six-hour-old barley seedlings were transferred to 100 μg/ ml solution of bleomycin A5 hydrochloride (LKT Laboratories) and incubated at 37°C for 2 h. Upon the mutagenic treatment the seedlings were washed three times in sterile water for 15 min.

Techniques: Control

Journal: Biosensors

Article Title: Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression

doi: 10.3390/bios2030318

Figure Lengend Snippet: The cytotoxicity of bleomycin, mitomycin C and ethanol to flounder gill (FG) cells as determined by thiazolyl blue tetrazolium bromide ( MTT) assay. Data are expressed as mean ± SD.

Article Snippet: Bleomycin sulfate, Mitomycin C, Geneticin (G418), Lipofectamine LTX and PLUS reagents were purchased from Invitrogen, USA.

Techniques: MTT Assay

Journal: Biosensors

Article Title: Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression

doi: 10.3390/bios2030318

Figure Lengend Snippet: Validation of the endogenous p53 -signaling pathway in FG cells. Examination of the responses of the transiently transformed FG cells to genotoxicant of bleomycin (30 μg/mL for 4 h) using firefly luciferase reporter plasmids of pGL 3 -p21-luc and pGL 3 -p53-luc and the Renilla luciferase internal reference plasmid of pRL-CMV. The intact FG cells (not transformed) were used as control. Data are expressed as mean ± SD.

Article Snippet: Bleomycin sulfate, Mitomycin C, Geneticin (G418), Lipofectamine LTX and PLUS reagents were purchased from Invitrogen, USA.

Techniques: Biomarker Discovery, Transformation Assay, Luciferase, Plasmid Preparation, Control

Journal: Biosensors

Article Title: Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression

doi: 10.3390/bios2030318

Figure Lengend Snippet: The time-course responses of the stable p21FGLuc cells to model genotoxic and non-genotoxic agents. The stable p21FGLuc cells were exposed to 30 μg /mL bleomycin, 10 μg /mL mitomycin C and 30% (v/v) ethanol, respectively. The same stable p21FGLuc cells, not exposed to any toxicants but with the same volume of PBS, were used as control. ** Shows the highly significant difference (p < 0.01). Data are expressed as mean ± SD.

Article Snippet: Bleomycin sulfate, Mitomycin C, Geneticin (G418), Lipofectamine LTX and PLUS reagents were purchased from Invitrogen, USA.

Techniques: Control

Journal: Biosensors

Article Title: Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression

doi: 10.3390/bios2030318

Figure Lengend Snippet: The dose-dependent responses of the stable p21FGLuc cells to model genotoxic and non-genotoxic agents. The stable p21FGLuc cells were exposed to increasing concentrations of bleomycin, mytomycin C and ethanol. Data are expressed as mean ± SD.

Article Snippet: Bleomycin sulfate, Mitomycin C, Geneticin (G418), Lipofectamine LTX and PLUS reagents were purchased from Invitrogen, USA.

Techniques:

Journal: Biosensors

Article Title: Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression

doi: 10.3390/bios2030318

Figure Lengend Snippet: Comparison of the DNA damage-induced responses of the stable p21FGLuc cells to bleomycin, mitomycin C and ethanol. The p21FGLuc cells were exposed to 30 μg/mL bleomycin for 4 h, 10 μg /mL mitomycin C for 2 h and 30% (v/v) ethanol for 1 h, respectively. ** Shows the highly significant difference (p < 0.01). *** Shows the very highly significant difference (p < 0.001). Data are expressed as mean ± SD.

Article Snippet: Bleomycin sulfate, Mitomycin C, Geneticin (G418), Lipofectamine LTX and PLUS reagents were purchased from Invitrogen, USA.

Techniques: Comparison

Journal: Biosensors

Article Title: Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression

doi: 10.3390/bios2030318

Figure Lengend Snippet: Summary of the genotoxicity detection results using the fish cell biosensor system (p21FGLuc).

Article Snippet: Bleomycin sulfate, Mitomycin C, Geneticin (G418), Lipofectamine LTX and PLUS reagents were purchased from Invitrogen, USA.

Techniques: Concentration Assay

Journal: Data in Brief

Article Title: Data on plasma tumour and metabolism related proteins’ potential in differentiation of HFpEF-PH from PAH and in prognosis of left heart failure patients with pulmonary hypertension

doi: 10.1016/j.dib.2021.107747

Figure Lengend Snippet: Plasma levels of metabolism and tumour related proteins in controls and patients.

Article Snippet: NT-proBNP and 69 tumour and metabolism related proteins were analysed: 5′-nucleotidase (5′-NT), protein AMBP or alpha-1-microglobulin/bikunin precursor (AMBP), aminopeptidase N (AP-N), bleomycin hydrolase (BLM-H), brother of cell adhesion molecule-related/down-regulated by oncogenes (CDO) or (BOC), carbonic anhydrase 9 (CA9), cathepsin Z, cyclin-dependant kinase inhibitor 1A (CDKN1A) or p21, carcinoembryonic antigen-related cell adhesion molecule (CEACAM) 1 and 5, contactin-1, cornulin, carboxypeptidase (CP) A1, B1 and E, cystatin B, decorin, endothelial cell-specific molecule 1 or endocan, epithelial cell adhesion molecule (EpCAM), fatty acid-binding protein 4 (FABP4), fibroblast growth factor (FGF-) 21 and 23, folate receptor gamma (FR-gamma), furin, gastrotropin, glyoxalase I or lactoylglutathione lyase, insulin-like growth factor 1 receptor (IGF1R), insulin-like growth factor-binding protein (IGFBP) 2, 3 and 7, kallikrein 6, 8, 11, 13 and 14, low-density lipoprotein receptor (LDL-R), leptin, lectin-like oxidized LDL receptor 1 (LOX-1), lipoprotein lipase (LPL), Ly6/PLAUR domain-containing protein 3 (LYPD3) or C4.4A, mesothelin, methionine aminopeptidase 2 (MetAP2), melanoma-derived growth regulatory protein (MIA), midkine, mucin-16 or CA125, nectin-4 or PVRL4, pappalysin-1, proprotein convertase subtilisin/kexin type 9 (PCSK9), podocalyxin, paraoxonase-3 (PON-3), prostasin, protein S100A4 (S100A4), protein S100A11 (S100A11), retinoic acid receptor responder protein 2 (RARRES2), resistin, secretory carrier-associated membrane protein 3 (SCAMP3), secretoglobin family 3A member 2 (SCGB3A2), serpin A12, tyrosine-protein phosphatase non-receptor type substrate 1 (SHPS-1), sortillin, soluble receptor for advanced glycation end products (sRAGE; all soluble forms of RAGE), T-cell leukaemia/lymphoma protein 1A (TCL1A), trefoil factor 3 (TFF3), protein-glutamine gamma-glutamyltransferase 2 (TGM2), transferrin receptor protein 1 (TR), WAP four-disulfide core domain protein 2 (WFDC2), vimentin, V-set and immunoglobulin domain-containing protein 2 (VSIG2) and Xaa-Pro aminopeptidase 2 (XPNPEP2).

Techniques:

Journal: Data in Brief

Article Title: Data on plasma tumour and metabolism related proteins’ potential in differentiation of HFpEF-PH from PAH and in prognosis of left heart failure patients with pulmonary hypertension

doi: 10.1016/j.dib.2021.107747

Figure Lengend Snippet: Proteins’ classification and p-values of Kruskal Wallis and Mann Whitney's tests in comparing metabolism and tumour related proteins in controls and disease groups.

Article Snippet: NT-proBNP and 69 tumour and metabolism related proteins were analysed: 5′-nucleotidase (5′-NT), protein AMBP or alpha-1-microglobulin/bikunin precursor (AMBP), aminopeptidase N (AP-N), bleomycin hydrolase (BLM-H), brother of cell adhesion molecule-related/down-regulated by oncogenes (CDO) or (BOC), carbonic anhydrase 9 (CA9), cathepsin Z, cyclin-dependant kinase inhibitor 1A (CDKN1A) or p21, carcinoembryonic antigen-related cell adhesion molecule (CEACAM) 1 and 5, contactin-1, cornulin, carboxypeptidase (CP) A1, B1 and E, cystatin B, decorin, endothelial cell-specific molecule 1 or endocan, epithelial cell adhesion molecule (EpCAM), fatty acid-binding protein 4 (FABP4), fibroblast growth factor (FGF-) 21 and 23, folate receptor gamma (FR-gamma), furin, gastrotropin, glyoxalase I or lactoylglutathione lyase, insulin-like growth factor 1 receptor (IGF1R), insulin-like growth factor-binding protein (IGFBP) 2, 3 and 7, kallikrein 6, 8, 11, 13 and 14, low-density lipoprotein receptor (LDL-R), leptin, lectin-like oxidized LDL receptor 1 (LOX-1), lipoprotein lipase (LPL), Ly6/PLAUR domain-containing protein 3 (LYPD3) or C4.4A, mesothelin, methionine aminopeptidase 2 (MetAP2), melanoma-derived growth regulatory protein (MIA), midkine, mucin-16 or CA125, nectin-4 or PVRL4, pappalysin-1, proprotein convertase subtilisin/kexin type 9 (PCSK9), podocalyxin, paraoxonase-3 (PON-3), prostasin, protein S100A4 (S100A4), protein S100A11 (S100A11), retinoic acid receptor responder protein 2 (RARRES2), resistin, secretory carrier-associated membrane protein 3 (SCAMP3), secretoglobin family 3A member 2 (SCGB3A2), serpin A12, tyrosine-protein phosphatase non-receptor type substrate 1 (SHPS-1), sortillin, soluble receptor for advanced glycation end products (sRAGE; all soluble forms of RAGE), T-cell leukaemia/lymphoma protein 1A (TCL1A), trefoil factor 3 (TFF3), protein-glutamine gamma-glutamyltransferase 2 (TGM2), transferrin receptor protein 1 (TR), WAP four-disulfide core domain protein 2 (WFDC2), vimentin, V-set and immunoglobulin domain-containing protein 2 (VSIG2) and Xaa-Pro aminopeptidase 2 (XPNPEP2).

Techniques:

Journal: Immunobiology

Article Title: HMGB1 regulates the activation of dendritic cells and CD4+ T cell responses through the modulation of autophagy in bleomycin-induced pulmonary fibrosis.

doi: 10.1016/j.imbio.2025.152906

Figure Lengend Snippet: Fig. 1. Activation of dendritic cells in the serum and lung tissue of a bleomycin-induced pulmonary interstitial fibrosis mouse model. (A-B) flow cytometry dot plots showing the distribution of MHCII+/CD11c + cells in the lung tissue and peripheral blood of sham and model mice. (C–D) flow cytometry dot plots showing the distribution of CD80+/CD40+ cells in the lung tissue and peripheral blood of sham and model mice. (E-F) flow cytometry dot plots showing the distribution of CD86+ cells in the lung tissue and peripheral blood of sham and model mice. (G-L) the number of MHCII+/CD11c+, CD80+/CD40+, and CD86+ cells in the lung tissue and peripheral blood of sham and model mice is presented in bar charts. **P < 0.01 vs. Sham group.

Article Snippet: Model group: the indwelling needle was inserted down the trachea into the airway and complete the infusion of 1.5 mg/kg bleomycin (HY-K1053, MedChemExpress, Shanghai, China) to induce lung fibrosis.

Techniques: Activation Assay, Flow Cytometry

Journal: Immunobiology

Article Title: HMGB1 regulates the activation of dendritic cells and CD4+ T cell responses through the modulation of autophagy in bleomycin-induced pulmonary fibrosis.

doi: 10.1016/j.imbio.2025.152906

Figure Lengend Snippet: Fig. 2. High expression of HMGB1 and increased autophagy and inflammatory infiltration in the lung tissue of bleomycin-induced pulmonary interstitial fibrosis mouse model. (A-B) the expression of LC3B was detected by western blot. (C-E) the expression of TNF-α, IL-1β, and HMGB1 was detected by ELISA. (F) the path ological changes in lung tissue of Sham and model groups were detected by HE staining. **P < 0.01 vs. Sham group.

Article Snippet: Model group: the indwelling needle was inserted down the trachea into the airway and complete the infusion of 1.5 mg/kg bleomycin (HY-K1053, MedChemExpress, Shanghai, China) to induce lung fibrosis.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Staining

Journal: Immunobiology

Article Title: HMGB1 regulates the activation of dendritic cells and CD4+ T cell responses through the modulation of autophagy in bleomycin-induced pulmonary fibrosis.

doi: 10.1016/j.imbio.2025.152906

Figure Lengend Snippet: Fig. 5. HMGB1 promotes DCs activation through regulating autophagy. Rapamycin and 3-MA were used to treat DCs from a bleomycin-induced mice model of pulmonary interstitial fibrosis, followed by HMGB1 stimulation for activation. (A-C) flow cytometry scatter plots show the distribution of MHCI+/CD11c+, CD80+/ CD40+, and CD86+ cells in each treatment group. (D–F) the number of MHCI+/CD11c+, CD80+/CD40+, and CD86+ cells in each treatment group is presented through bar graphs. **P < 0.01 vs. Control group, ##P < 0.01 vs. HMGB1 group.

Article Snippet: Model group: the indwelling needle was inserted down the trachea into the airway and complete the infusion of 1.5 mg/kg bleomycin (HY-K1053, MedChemExpress, Shanghai, China) to induce lung fibrosis.

Techniques: Activation Assay, Flow Cytometry, Control