Etoposide (40 µM) was added to oocytes for 15 min before NEB, or to maturing oocytes for the times as indicated. 5-Iodotubercidin (Cambridge Bioscience, CAY10010375) was dissolved in 100% ethanol and diluted in media at 1:1000 to give a working concentration of 0.5 µM. Other additions were nocodazole (400 nM); ZM447439 (10 µM; Bio-Techne, MN, USA), Mps1 inhibitor AZ3146 (2 µM; Bio-Techne), MG132 (10 µM), KU55933 (10 µM; Merck-Millipore, UK), bleomycin (1 µM; Abcam, UK) and ATR kinase inhibitor II (10 µM; Merck-Millipore, UK). All drugs were dissolved in DMSO and supplemented with the neutral detergent 200 µg ml−1 pluronic acid to aid dispersion, and used at dilutions of 0.1% or below.
Bleomycin
Manufactured by Abcam
Sourced in United Kingdom
Bleomycin is a cytotoxic glycopeptide antibiotic used as a laboratory reagent. It is primarily employed in research applications to induce DNA damage and cell death.
Automatically generated - may contain errors
Lab products found in correlation
Mg132, by Merck Group (1 mentions)
Ku55933, by Merck Group (1 mentions)
Anti cd8a clone 53 6.7, by BioLegend (1 mentions)
Ionomycin, by Abcam (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Abcam (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Bio-Optica (1 mentions)
Tissue tek oct, by Thermo Fisher Scientific (1 mentions)
Fluoroshield with dapi, by Vector Laboratories (1 mentions)
Gelred nucleic acid, by Biotium (1 mentions)
Prestoblue cell viability reagent, by Thermo Fisher Scientific (1 mentions)
Masson s trichrome, by Bio-Optica (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Qpcrbio sygreen 1 step go lo rox, by PCR Biosystems (1 mentions)
4 protocols using bleomycin
1
Oocyte Maturation Modulation Protocols
2
Tumor-Infiltrating T Cell Analysis
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Tumors and lymph nodes of the mice in each group were collected 7 days after the last injection, digested and grinded to single cell suspension. Tumor cell suspension was harvested for tumor-infiltrating T cell (TIL) analysis. TILs were stained with antibodies (Biolegend, USA): anti-CD3 (clone 145-2C11), anti-CD4 (clone GK1.5), and anti-CD8a (clone 53-6.7) for 30 min. To detect intracellular cytokine IFN-γ, tumor cell suspension was incubated with ionomycin (1 mg mL−1; Abcam), phorbol 12-myristate 13-acetate (PMA; 50 ng mL−1, Abcam) and bleomycin (1 mg mL−1; Abcam) for 4–6 h. After that, cells were fixed, permeated and stained with anti-IFN-γ (clone XMG1.2). Popliteal and inguinal lymph node cell suspension were harvested to detect the maturation of DCs. DCs were stained with surface antibodies: anti-CD11c (clone N418), anti-CD80 (clone 16-10A1), and anti-CD86 (clone GL-1) for 30 min. Cells were then washed and resuspended in fresh PBS, and analyzed by flow cytometry. All flow cytometry antibodies were purchased from Biolegend.
3
Inducing Midgut Damage in Anopheles
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To induce chemical damage in the midgut, 3- to 4 day-old females of An. stephensi HP10 hindgut line were fed with 10% Karo syrup solution supplemented with 25 µg/ml Bleomycin (Bleomycin sulfate, anticancer and antibiotic agent, ab142977, Abcam, Cambridge, UK) for two days25 (link). The fresh solution was offered every day of the treatment. After two days, females were artificially fed with 10% BSA saline solution to distend the midgut tissue as described above.
4
Multimodal Analysis of HCC1954 Cell Line
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PCL (MW 60 kDa), sodium cacodylate, sucrose, acetonitrile, propidium iodide (PI), Hoechst 33342 (HOE), hexafluoroisopropanol (HFIP), doxorubicin, and bovine serum albumin (BSA) were obtained from Sigma Aldrich (Steinheim, Germany), and ethanol from VWR Chemicals Prolab (Fontenay-sous-Bois, France). Bleomycin was from Abcam (Cambridge, UK).
The HCC1954 cell line was provided by ATTC (Guernsey, UK). Fetal bovine serum (FBS), culture media, and supplements were obtained from Corning (Mediatech Inc., Manassas, VA, USA). The PrestoBlue™ Cell Viability Reagent and secondary antibody goat anti-mouse Alexa 488 were purchased by Thermo Fisher Scientific (Eugene, OR, USA), whereas paraformaldehyde, Masson’s trichrome, and Alcian blue staining kits by BioOptica (Milan, Italy). The TriReagent and the qPCRBIO SyGreen 1-Step Go Lo-ROX were purchased from PCRBIOSYSTEMS (London, UK), and Gel Red Nucleic Acid staining from Biotium (Hayward, CA, USA). Anti-CD44 antibody was provided by AbD Serotech (Kidlington, UK), and Fluoroshield with DAPI was from Vector Laboratories (Peterborough, UK). Tissue-Tek® OCT was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
The HCC1954 cell line was provided by ATTC (Guernsey, UK). Fetal bovine serum (FBS), culture media, and supplements were obtained from Corning (Mediatech Inc., Manassas, VA, USA). The PrestoBlue™ Cell Viability Reagent and secondary antibody goat anti-mouse Alexa 488 were purchased by Thermo Fisher Scientific (Eugene, OR, USA), whereas paraformaldehyde, Masson’s trichrome, and Alcian blue staining kits by BioOptica (Milan, Italy). The TriReagent and the qPCRBIO SyGreen 1-Step Go Lo-ROX were purchased from PCRBIOSYSTEMS (London, UK), and Gel Red Nucleic Acid staining from Biotium (Hayward, CA, USA). Anti-CD44 antibody was provided by AbD Serotech (Kidlington, UK), and Fluoroshield with DAPI was from Vector Laboratories (Peterborough, UK). Tissue-Tek® OCT was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
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