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pcmv spike  (Addgene inc)


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    Addgene inc pcmv spike
    Pcmv Spike, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/plenti+cmv+to+neo+dest/pLenti+CMV+Neo+DEST+(705-1)+(Plasmid+%2317392)/pm41430043-261-11-12
    Average 93 stars, based on 60 article reviews
    pcmv spike - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Over Expression:

    Article Title: Oncogene-like induction of cellular invasion from centrosome amplification.
    Article Snippet: .. For the generation of the inducible PLK4 overexpression system we used the lentiviral vectors pLenti-CMV-TetR-Blast (17492, Addgene) and pLenti-CMV/TO-Neo-Dest (17292, Addgene)39. .. Wild-type PLK4 and PLK41–608 cDNAs were cloned using the Gateway system into the pLentiCMV/TO-Neo-Dest vector.

    Article Title: PARP inhibitors suppress tumours via centrosome error-induced senescence independent of DNA damage response
    Article Snippet: .. For inducible PLK4 overexpression in MCF7 cells, the human PLK4 coding sequence was subcloned into pLenti-CMV/TO-Neo-Dest (Addgene, Cat# 17292). .. For Flag-BRCA1 or Flag-ZNF721 overexpression in MCF7 cells, the human BRCA1 or ZNF721 coding sequence was subcloned into pcDNA3.1-Flag (Addgene, Cat# 208051).

    Article Title: PARP inhibitors suppress tumours via centrosome error-induced senescence independent of DNA damage response.
    Article Snippet: .. For inducible PLK4 overexpression in MCF7 cells, the human PLK4 coding sequence was subcloned into pLenti-CMV/TO-Neo-Dest (Addgene, Cat# 17292). .. For Flag-BRCA1 or Flag-ZNF721 overexpression in MCF7 cells, the human BRCA1 or ZNF721 coding sequence was subcloned into pcDNA3.1-Flag (Addgene, Cat# 208051).

    Clone Assay:

    Article Title: Investigation of the role of tyrosine kinase receptor EPHA3 in colorectal cancer
    Article Snippet: LS174T ( EPHA3 wild type) and DLD1 (heterozygous EPHA3 T719I and M847K mutations) cells carrying the tet-repressor plasmid (TR1 and TR7, respectively) were generated using the T-rex system (Invitrogen) as described previously . .. To generate EPHA3-inducible clones, DLD1-TR7 and LS174T-TR1 expressing the Tet repressor were transfected with pLenti-CMV/TO-EPHA3 and the corresponding empty vector (pLenti-CMV/TO Neo DEST, Addgene #17292) with Lipofectamine 2000. .. Transfectants were selected in medium containing G418 (1 or 0.5 mg/ml for DLD1 and LS174T respectively; Invitrogen).

    Expressing:

    Article Title: Investigation of the role of tyrosine kinase receptor EPHA3 in colorectal cancer
    Article Snippet: LS174T ( EPHA3 wild type) and DLD1 (heterozygous EPHA3 T719I and M847K mutations) cells carrying the tet-repressor plasmid (TR1 and TR7, respectively) were generated using the T-rex system (Invitrogen) as described previously . .. To generate EPHA3-inducible clones, DLD1-TR7 and LS174T-TR1 expressing the Tet repressor were transfected with pLenti-CMV/TO-EPHA3 and the corresponding empty vector (pLenti-CMV/TO Neo DEST, Addgene #17292) with Lipofectamine 2000. .. Transfectants were selected in medium containing G418 (1 or 0.5 mg/ml for DLD1 and LS174T respectively; Invitrogen).

    Transfection:

    Article Title: Investigation of the role of tyrosine kinase receptor EPHA3 in colorectal cancer
    Article Snippet: LS174T ( EPHA3 wild type) and DLD1 (heterozygous EPHA3 T719I and M847K mutations) cells carrying the tet-repressor plasmid (TR1 and TR7, respectively) were generated using the T-rex system (Invitrogen) as described previously . .. To generate EPHA3-inducible clones, DLD1-TR7 and LS174T-TR1 expressing the Tet repressor were transfected with pLenti-CMV/TO-EPHA3 and the corresponding empty vector (pLenti-CMV/TO Neo DEST, Addgene #17292) with Lipofectamine 2000. .. Transfectants were selected in medium containing G418 (1 or 0.5 mg/ml for DLD1 and LS174T respectively; Invitrogen).

    Bradford Protein Assay:

    Article Title: Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Alexa-conjugated A488 Life Technologies Cat#A11008; RRID: AB_143165 Mouse Alexa-conjugated A568 Life Technologies Cat#A11001, RRID: AB_2534069 Rabbit Alexa-conjugated A568 Life Technologies Cat#A11011, RRID: AB_143157 Mouse a-tubulin Sigma-Aldrich Cat#T9026; RRID: AB_477593 Rabbit centrin2 N-17-R Santa Cruz Cat#sc-27793-R; RRID: AB_2082359 Mouse LBPA (6C4) Merck Millipore Cat#MABT837 Rabbit LC3B (D11) XP Cell signaling Cat#3868S; RRID: AB_2137707 Mouse a-SMA Sigma-Aldrich Cat#A2547; RRID: AB_476701 Mouse Ki67 Alexa-conjugated A488 BD Biosciences Cat#561165; RRID: AB_10611866 Rabbit TSG101 [(EPR7130(b)] Abcam Cat#ab125011; RRID: AB_10974262 Rabbit CD63 Abcam Cat#ab68418; RRID: AB_10563972 Mouse CD81 (clone B-11) Santa Cruz Biotechnology Cat#sc-166029; RRID: AB_2275892 Mouse ALIX (clone 3A9) Cell signaling Cat#2171, RRID: AB_2299455 Mouse Flotillin-1 (clone 18) Biosciences Cat#610821; RRID: AB_398140 HRP- anti rabbit secondary GE Healthcare Cat#NA934; RRID: AB_772206 HRP- anti mouse secondary GE Healthcare Cat#NA931; RRID: AB_772210 Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma-Aldrich #D9891 H2O2 Sigma-Aldrich #H1009 N-acetyl cysteine Sigma-Aldrich #A9165 Bafilomycin A1 Sigma-Aldrich #B1793-10UG DMEM/F12 Sigma-Aldrich #D8437 DMEM Thermo Fisher Scientific #41966-029 RPMI-1640 Thermo Fisher Scientific #11875093 Keratinocyte serum free medium (1X) Thermo Fisher Scientific #17005042 Opti-MEM reduced serum medium Thermo Fisher Scientific #31985070 DMEM for SILAC Thermo Fisher Scientific #88364 Penicillin/Streptomycin Thermo Fisher Scientific #15140-122 FBS Thermo Fisher Scientific #10500-064 Tet-free FBS Hyclone #SH30070.03T GIBCO FBS, Dialyzed Thermo Fisher Scientific #26400044 Blasticidin Generon #2805-10 Geneticin (G418) Thermo Fisher Scientific #10131027 Puromycin InvivoGen #ant-pr-1 Polybrene Sigma-Aldrich #H9268 Formaldehyde 16% Thermo Fisher Scientific #28908 Alexa Fluor 568 Phalloidin Life Technologies #A12380 Hoechst 33342 ThermoFisher Scientific # H3570 BODIPY FL N-(2-Aminoethyl) Maleimide Thermo Fisher Scientific #B10250 ProLong anti-fade mounting medium Molecular Probes #P36934 BSA Sigma-Aldrich #A9647 Lipofectamine 2000 Invitrogen #11668027 Lipofectamine RNAi Max Invitrogen #13778075 RIPA Buffer Thermo Scientific #89901 Complete Mini Protease Inhibitor Cocktail Roche #11836153001 (Continued on next page) Current Biology 31, 1403–1416.e1–e7, April 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phosphatase inhibitor Cocktail Cell Signaling #5870 Bradford Protein Assay Bio-Rad #5000006 Recombinant Human TGF-b1 PEPROTECH #100-21 Matrigel Matrix Basement Membrane Life Sciences #354234 Corning Rat Tail High Concentration Life Sciences #354249 DTT VWR Chemicals #M109; CAS: 3483-12-3 Iodoacetamide VWR Chemicals Cat#786-228; CAS: 144-48-9 Trypsin Sigma-Aldrich T6567-1MG L-Arginine Sigma-Aldrich A6969-25G L-Lysine Sigma-Aldrich L8662-25G L-Arginine [U-13C6] Cambridge Isotopes CLM-2265-H-0.5 L-Lysine [4,4,5,5-D4] Cambridge Isotopes DLM-2640-0.5 L-Arginine [U-13C6, U-15N4] Cambridge Isotopes CNLM-539-H-0.5 L-Lysine [U-13C6, U-15N2] Cambridge Isotopes CNLM-291-H-0.5 L-Proline Sigma-Aldrich P0380-100G Critical commercial assays GSH/GSSH-Glo assay Promega #V6611 BCA Protein assay Thermo Fisher Scientific #23225 Magic Red Cathepsin B kit Bio-Rad #ICT937 CellVue Maroon Cell Labeling Kit Invitrogen #88-0870-16 Deposited data SILAC analysis of exosome secretion in response to centrosome amplification This paper PRIDE: PXD020984 accessible via PRIDE partner repository (https://www.ebi. ac.uk/pride/archive/) Experimental models: cell lines PaTu-8988T Prof. Hemant Kocher (QMUL) N/A PaTu-8988S Prof. Yaohe Wang (QMUL) N/A Panc-1 Prof. Hemant Kocher (QMUL) N/A CFPAC-1 Prof. David Pellman (Harvard) N/A Capan-1 Prof. Hemant Kocher (QMUL) N/A HPAF-II Prof. Hemant Kocher (QMUL) N/A MIA-PaCa-2 Prof. Hemant Kocher (QMUL) N/A DEC-hTERT Prof. Hemant Kocher (QMUL) N/A HPDE Prof. Yaohe Wang (QMUL) N/A PS1 Prof. Hemant Kocher (QMUL) N/A HEK293M Prof. David Pellman (Harvard) N/A PaTu-8988S.PLK4 This work N/A HPAF-II.PLK4 This work N/A HPAF-II.PLK4 H2B RFP This work N/A PS1 H2B GFP Angus Cameron (QMUL) N/A Oligonucleotides siSAS6 on-TARGET smart pool Dharmacon #M-004158-02 siNegative Control QIAGEN #1027310 Recombinant DNA pLenti-CMV-TetR-Blast Addgene #17492 pLenti-CMV/TO-Neo-DEST.PLK4 25 N/A pMD2.G VSV-G Addgene #12259 psPAX2 Gag-Pol Addgene #12260 (Continued on next page) ll OPEN ACCESS e2 Current Biology 31, 1403–1416.e1–e7, April 12, 2021 Article .. REAGENT or RESOURCE SOURCE IDENTIFIER Other qEV original Size Exclusion Chromatography (SEC) columns izon #SP1 Tube, Thinwall, Ultra-Clear, 38.5 mL, 25 3 89 mm Beckman coulter #344058 Amicon Ultra-15 Centrifugal Filter Unit Merk #UFC9010 ll OPEN ACCESSArticle

    Recombinant:

    Article Title: Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Alexa-conjugated A488 Life Technologies Cat#A11008; RRID: AB_143165 Mouse Alexa-conjugated A568 Life Technologies Cat#A11001, RRID: AB_2534069 Rabbit Alexa-conjugated A568 Life Technologies Cat#A11011, RRID: AB_143157 Mouse a-tubulin Sigma-Aldrich Cat#T9026; RRID: AB_477593 Rabbit centrin2 N-17-R Santa Cruz Cat#sc-27793-R; RRID: AB_2082359 Mouse LBPA (6C4) Merck Millipore Cat#MABT837 Rabbit LC3B (D11) XP Cell signaling Cat#3868S; RRID: AB_2137707 Mouse a-SMA Sigma-Aldrich Cat#A2547; RRID: AB_476701 Mouse Ki67 Alexa-conjugated A488 BD Biosciences Cat#561165; RRID: AB_10611866 Rabbit TSG101 [(EPR7130(b)] Abcam Cat#ab125011; RRID: AB_10974262 Rabbit CD63 Abcam Cat#ab68418; RRID: AB_10563972 Mouse CD81 (clone B-11) Santa Cruz Biotechnology Cat#sc-166029; RRID: AB_2275892 Mouse ALIX (clone 3A9) Cell signaling Cat#2171, RRID: AB_2299455 Mouse Flotillin-1 (clone 18) Biosciences Cat#610821; RRID: AB_398140 HRP- anti rabbit secondary GE Healthcare Cat#NA934; RRID: AB_772206 HRP- anti mouse secondary GE Healthcare Cat#NA931; RRID: AB_772210 Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma-Aldrich #D9891 H2O2 Sigma-Aldrich #H1009 N-acetyl cysteine Sigma-Aldrich #A9165 Bafilomycin A1 Sigma-Aldrich #B1793-10UG DMEM/F12 Sigma-Aldrich #D8437 DMEM Thermo Fisher Scientific #41966-029 RPMI-1640 Thermo Fisher Scientific #11875093 Keratinocyte serum free medium (1X) Thermo Fisher Scientific #17005042 Opti-MEM reduced serum medium Thermo Fisher Scientific #31985070 DMEM for SILAC Thermo Fisher Scientific #88364 Penicillin/Streptomycin Thermo Fisher Scientific #15140-122 FBS Thermo Fisher Scientific #10500-064 Tet-free FBS Hyclone #SH30070.03T GIBCO FBS, Dialyzed Thermo Fisher Scientific #26400044 Blasticidin Generon #2805-10 Geneticin (G418) Thermo Fisher Scientific #10131027 Puromycin InvivoGen #ant-pr-1 Polybrene Sigma-Aldrich #H9268 Formaldehyde 16% Thermo Fisher Scientific #28908 Alexa Fluor 568 Phalloidin Life Technologies #A12380 Hoechst 33342 ThermoFisher Scientific # H3570 BODIPY FL N-(2-Aminoethyl) Maleimide Thermo Fisher Scientific #B10250 ProLong anti-fade mounting medium Molecular Probes #P36934 BSA Sigma-Aldrich #A9647 Lipofectamine 2000 Invitrogen #11668027 Lipofectamine RNAi Max Invitrogen #13778075 RIPA Buffer Thermo Scientific #89901 Complete Mini Protease Inhibitor Cocktail Roche #11836153001 (Continued on next page) Current Biology 31, 1403–1416.e1–e7, April 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phosphatase inhibitor Cocktail Cell Signaling #5870 Bradford Protein Assay Bio-Rad #5000006 Recombinant Human TGF-b1 PEPROTECH #100-21 Matrigel Matrix Basement Membrane Life Sciences #354234 Corning Rat Tail High Concentration Life Sciences #354249 DTT VWR Chemicals #M109; CAS: 3483-12-3 Iodoacetamide VWR Chemicals Cat#786-228; CAS: 144-48-9 Trypsin Sigma-Aldrich T6567-1MG L-Arginine Sigma-Aldrich A6969-25G L-Lysine Sigma-Aldrich L8662-25G L-Arginine [U-13C6] Cambridge Isotopes CLM-2265-H-0.5 L-Lysine [4,4,5,5-D4] Cambridge Isotopes DLM-2640-0.5 L-Arginine [U-13C6, U-15N4] Cambridge Isotopes CNLM-539-H-0.5 L-Lysine [U-13C6, U-15N2] Cambridge Isotopes CNLM-291-H-0.5 L-Proline Sigma-Aldrich P0380-100G Critical commercial assays GSH/GSSH-Glo assay Promega #V6611 BCA Protein assay Thermo Fisher Scientific #23225 Magic Red Cathepsin B kit Bio-Rad #ICT937 CellVue Maroon Cell Labeling Kit Invitrogen #88-0870-16 Deposited data SILAC analysis of exosome secretion in response to centrosome amplification This paper PRIDE: PXD020984 accessible via PRIDE partner repository (https://www.ebi. ac.uk/pride/archive/) Experimental models: cell lines PaTu-8988T Prof. Hemant Kocher (QMUL) N/A PaTu-8988S Prof. Yaohe Wang (QMUL) N/A Panc-1 Prof. Hemant Kocher (QMUL) N/A CFPAC-1 Prof. David Pellman (Harvard) N/A Capan-1 Prof. Hemant Kocher (QMUL) N/A HPAF-II Prof. Hemant Kocher (QMUL) N/A MIA-PaCa-2 Prof. Hemant Kocher (QMUL) N/A DEC-hTERT Prof. Hemant Kocher (QMUL) N/A HPDE Prof. Yaohe Wang (QMUL) N/A PS1 Prof. Hemant Kocher (QMUL) N/A HEK293M Prof. David Pellman (Harvard) N/A PaTu-8988S.PLK4 This work N/A HPAF-II.PLK4 This work N/A HPAF-II.PLK4 H2B RFP This work N/A PS1 H2B GFP Angus Cameron (QMUL) N/A Oligonucleotides siSAS6 on-TARGET smart pool Dharmacon #M-004158-02 siNegative Control QIAGEN #1027310 Recombinant DNA pLenti-CMV-TetR-Blast Addgene #17492 pLenti-CMV/TO-Neo-DEST.PLK4 25 N/A pMD2.G VSV-G Addgene #12259 psPAX2 Gag-Pol Addgene #12260 (Continued on next page) ll OPEN ACCESS e2 Current Biology 31, 1403–1416.e1–e7, April 12, 2021 Article .. REAGENT or RESOURCE SOURCE IDENTIFIER Other qEV original Size Exclusion Chromatography (SEC) columns izon #SP1 Tube, Thinwall, Ultra-Clear, 38.5 mL, 25 3 89 mm Beckman coulter #344058 Amicon Ultra-15 Centrifugal Filter Unit Merk #UFC9010 ll OPEN ACCESSArticle

    Membrane:

    Article Title: Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Alexa-conjugated A488 Life Technologies Cat#A11008; RRID: AB_143165 Mouse Alexa-conjugated A568 Life Technologies Cat#A11001, RRID: AB_2534069 Rabbit Alexa-conjugated A568 Life Technologies Cat#A11011, RRID: AB_143157 Mouse a-tubulin Sigma-Aldrich Cat#T9026; RRID: AB_477593 Rabbit centrin2 N-17-R Santa Cruz Cat#sc-27793-R; RRID: AB_2082359 Mouse LBPA (6C4) Merck Millipore Cat#MABT837 Rabbit LC3B (D11) XP Cell signaling Cat#3868S; RRID: AB_2137707 Mouse a-SMA Sigma-Aldrich Cat#A2547; RRID: AB_476701 Mouse Ki67 Alexa-conjugated A488 BD Biosciences Cat#561165; RRID: AB_10611866 Rabbit TSG101 [(EPR7130(b)] Abcam Cat#ab125011; RRID: AB_10974262 Rabbit CD63 Abcam Cat#ab68418; RRID: AB_10563972 Mouse CD81 (clone B-11) Santa Cruz Biotechnology Cat#sc-166029; RRID: AB_2275892 Mouse ALIX (clone 3A9) Cell signaling Cat#2171, RRID: AB_2299455 Mouse Flotillin-1 (clone 18) Biosciences Cat#610821; RRID: AB_398140 HRP- anti rabbit secondary GE Healthcare Cat#NA934; RRID: AB_772206 HRP- anti mouse secondary GE Healthcare Cat#NA931; RRID: AB_772210 Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma-Aldrich #D9891 H2O2 Sigma-Aldrich #H1009 N-acetyl cysteine Sigma-Aldrich #A9165 Bafilomycin A1 Sigma-Aldrich #B1793-10UG DMEM/F12 Sigma-Aldrich #D8437 DMEM Thermo Fisher Scientific #41966-029 RPMI-1640 Thermo Fisher Scientific #11875093 Keratinocyte serum free medium (1X) Thermo Fisher Scientific #17005042 Opti-MEM reduced serum medium Thermo Fisher Scientific #31985070 DMEM for SILAC Thermo Fisher Scientific #88364 Penicillin/Streptomycin Thermo Fisher Scientific #15140-122 FBS Thermo Fisher Scientific #10500-064 Tet-free FBS Hyclone #SH30070.03T GIBCO FBS, Dialyzed Thermo Fisher Scientific #26400044 Blasticidin Generon #2805-10 Geneticin (G418) Thermo Fisher Scientific #10131027 Puromycin InvivoGen #ant-pr-1 Polybrene Sigma-Aldrich #H9268 Formaldehyde 16% Thermo Fisher Scientific #28908 Alexa Fluor 568 Phalloidin Life Technologies #A12380 Hoechst 33342 ThermoFisher Scientific # H3570 BODIPY FL N-(2-Aminoethyl) Maleimide Thermo Fisher Scientific #B10250 ProLong anti-fade mounting medium Molecular Probes #P36934 BSA Sigma-Aldrich #A9647 Lipofectamine 2000 Invitrogen #11668027 Lipofectamine RNAi Max Invitrogen #13778075 RIPA Buffer Thermo Scientific #89901 Complete Mini Protease Inhibitor Cocktail Roche #11836153001 (Continued on next page) Current Biology 31, 1403–1416.e1–e7, April 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phosphatase inhibitor Cocktail Cell Signaling #5870 Bradford Protein Assay Bio-Rad #5000006 Recombinant Human TGF-b1 PEPROTECH #100-21 Matrigel Matrix Basement Membrane Life Sciences #354234 Corning Rat Tail High Concentration Life Sciences #354249 DTT VWR Chemicals #M109; CAS: 3483-12-3 Iodoacetamide VWR Chemicals Cat#786-228; CAS: 144-48-9 Trypsin Sigma-Aldrich T6567-1MG L-Arginine Sigma-Aldrich A6969-25G L-Lysine Sigma-Aldrich L8662-25G L-Arginine [U-13C6] Cambridge Isotopes CLM-2265-H-0.5 L-Lysine [4,4,5,5-D4] Cambridge Isotopes DLM-2640-0.5 L-Arginine [U-13C6, U-15N4] Cambridge Isotopes CNLM-539-H-0.5 L-Lysine [U-13C6, U-15N2] Cambridge Isotopes CNLM-291-H-0.5 L-Proline Sigma-Aldrich P0380-100G Critical commercial assays GSH/GSSH-Glo assay Promega #V6611 BCA Protein assay Thermo Fisher Scientific #23225 Magic Red Cathepsin B kit Bio-Rad #ICT937 CellVue Maroon Cell Labeling Kit Invitrogen #88-0870-16 Deposited data SILAC analysis of exosome secretion in response to centrosome amplification This paper PRIDE: PXD020984 accessible via PRIDE partner repository (https://www.ebi. ac.uk/pride/archive/) Experimental models: cell lines PaTu-8988T Prof. Hemant Kocher (QMUL) N/A PaTu-8988S Prof. Yaohe Wang (QMUL) N/A Panc-1 Prof. Hemant Kocher (QMUL) N/A CFPAC-1 Prof. David Pellman (Harvard) N/A Capan-1 Prof. Hemant Kocher (QMUL) N/A HPAF-II Prof. Hemant Kocher (QMUL) N/A MIA-PaCa-2 Prof. Hemant Kocher (QMUL) N/A DEC-hTERT Prof. Hemant Kocher (QMUL) N/A HPDE Prof. Yaohe Wang (QMUL) N/A PS1 Prof. Hemant Kocher (QMUL) N/A HEK293M Prof. David Pellman (Harvard) N/A PaTu-8988S.PLK4 This work N/A HPAF-II.PLK4 This work N/A HPAF-II.PLK4 H2B RFP This work N/A PS1 H2B GFP Angus Cameron (QMUL) N/A Oligonucleotides siSAS6 on-TARGET smart pool Dharmacon #M-004158-02 siNegative Control QIAGEN #1027310 Recombinant DNA pLenti-CMV-TetR-Blast Addgene #17492 pLenti-CMV/TO-Neo-DEST.PLK4 25 N/A pMD2.G VSV-G Addgene #12259 psPAX2 Gag-Pol Addgene #12260 (Continued on next page) ll OPEN ACCESS e2 Current Biology 31, 1403–1416.e1–e7, April 12, 2021 Article .. REAGENT or RESOURCE SOURCE IDENTIFIER Other qEV original Size Exclusion Chromatography (SEC) columns izon #SP1 Tube, Thinwall, Ultra-Clear, 38.5 mL, 25 3 89 mm Beckman coulter #344058 Amicon Ultra-15 Centrifugal Filter Unit Merk #UFC9010 ll OPEN ACCESSArticle

    Concentration Assay:

    Article Title: Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Alexa-conjugated A488 Life Technologies Cat#A11008; RRID: AB_143165 Mouse Alexa-conjugated A568 Life Technologies Cat#A11001, RRID: AB_2534069 Rabbit Alexa-conjugated A568 Life Technologies Cat#A11011, RRID: AB_143157 Mouse a-tubulin Sigma-Aldrich Cat#T9026; RRID: AB_477593 Rabbit centrin2 N-17-R Santa Cruz Cat#sc-27793-R; RRID: AB_2082359 Mouse LBPA (6C4) Merck Millipore Cat#MABT837 Rabbit LC3B (D11) XP Cell signaling Cat#3868S; RRID: AB_2137707 Mouse a-SMA Sigma-Aldrich Cat#A2547; RRID: AB_476701 Mouse Ki67 Alexa-conjugated A488 BD Biosciences Cat#561165; RRID: AB_10611866 Rabbit TSG101 [(EPR7130(b)] Abcam Cat#ab125011; RRID: AB_10974262 Rabbit CD63 Abcam Cat#ab68418; RRID: AB_10563972 Mouse CD81 (clone B-11) Santa Cruz Biotechnology Cat#sc-166029; RRID: AB_2275892 Mouse ALIX (clone 3A9) Cell signaling Cat#2171, RRID: AB_2299455 Mouse Flotillin-1 (clone 18) Biosciences Cat#610821; RRID: AB_398140 HRP- anti rabbit secondary GE Healthcare Cat#NA934; RRID: AB_772206 HRP- anti mouse secondary GE Healthcare Cat#NA931; RRID: AB_772210 Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma-Aldrich #D9891 H2O2 Sigma-Aldrich #H1009 N-acetyl cysteine Sigma-Aldrich #A9165 Bafilomycin A1 Sigma-Aldrich #B1793-10UG DMEM/F12 Sigma-Aldrich #D8437 DMEM Thermo Fisher Scientific #41966-029 RPMI-1640 Thermo Fisher Scientific #11875093 Keratinocyte serum free medium (1X) Thermo Fisher Scientific #17005042 Opti-MEM reduced serum medium Thermo Fisher Scientific #31985070 DMEM for SILAC Thermo Fisher Scientific #88364 Penicillin/Streptomycin Thermo Fisher Scientific #15140-122 FBS Thermo Fisher Scientific #10500-064 Tet-free FBS Hyclone #SH30070.03T GIBCO FBS, Dialyzed Thermo Fisher Scientific #26400044 Blasticidin Generon #2805-10 Geneticin (G418) Thermo Fisher Scientific #10131027 Puromycin InvivoGen #ant-pr-1 Polybrene Sigma-Aldrich #H9268 Formaldehyde 16% Thermo Fisher Scientific #28908 Alexa Fluor 568 Phalloidin Life Technologies #A12380 Hoechst 33342 ThermoFisher Scientific # H3570 BODIPY FL N-(2-Aminoethyl) Maleimide Thermo Fisher Scientific #B10250 ProLong anti-fade mounting medium Molecular Probes #P36934 BSA Sigma-Aldrich #A9647 Lipofectamine 2000 Invitrogen #11668027 Lipofectamine RNAi Max Invitrogen #13778075 RIPA Buffer Thermo Scientific #89901 Complete Mini Protease Inhibitor Cocktail Roche #11836153001 (Continued on next page) Current Biology 31, 1403–1416.e1–e7, April 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phosphatase inhibitor Cocktail Cell Signaling #5870 Bradford Protein Assay Bio-Rad #5000006 Recombinant Human TGF-b1 PEPROTECH #100-21 Matrigel Matrix Basement Membrane Life Sciences #354234 Corning Rat Tail High Concentration Life Sciences #354249 DTT VWR Chemicals #M109; CAS: 3483-12-3 Iodoacetamide VWR Chemicals Cat#786-228; CAS: 144-48-9 Trypsin Sigma-Aldrich T6567-1MG L-Arginine Sigma-Aldrich A6969-25G L-Lysine Sigma-Aldrich L8662-25G L-Arginine [U-13C6] Cambridge Isotopes CLM-2265-H-0.5 L-Lysine [4,4,5,5-D4] Cambridge Isotopes DLM-2640-0.5 L-Arginine [U-13C6, U-15N4] Cambridge Isotopes CNLM-539-H-0.5 L-Lysine [U-13C6, U-15N2] Cambridge Isotopes CNLM-291-H-0.5 L-Proline Sigma-Aldrich P0380-100G Critical commercial assays GSH/GSSH-Glo assay Promega #V6611 BCA Protein assay Thermo Fisher Scientific #23225 Magic Red Cathepsin B kit Bio-Rad #ICT937 CellVue Maroon Cell Labeling Kit Invitrogen #88-0870-16 Deposited data SILAC analysis of exosome secretion in response to centrosome amplification This paper PRIDE: PXD020984 accessible via PRIDE partner repository (https://www.ebi. ac.uk/pride/archive/) Experimental models: cell lines PaTu-8988T Prof. Hemant Kocher (QMUL) N/A PaTu-8988S Prof. Yaohe Wang (QMUL) N/A Panc-1 Prof. Hemant Kocher (QMUL) N/A CFPAC-1 Prof. David Pellman (Harvard) N/A Capan-1 Prof. Hemant Kocher (QMUL) N/A HPAF-II Prof. Hemant Kocher (QMUL) N/A MIA-PaCa-2 Prof. Hemant Kocher (QMUL) N/A DEC-hTERT Prof. Hemant Kocher (QMUL) N/A HPDE Prof. Yaohe Wang (QMUL) N/A PS1 Prof. Hemant Kocher (QMUL) N/A HEK293M Prof. David Pellman (Harvard) N/A PaTu-8988S.PLK4 This work N/A HPAF-II.PLK4 This work N/A HPAF-II.PLK4 H2B RFP This work N/A PS1 H2B GFP Angus Cameron (QMUL) N/A Oligonucleotides siSAS6 on-TARGET smart pool Dharmacon #M-004158-02 siNegative Control QIAGEN #1027310 Recombinant DNA pLenti-CMV-TetR-Blast Addgene #17492 pLenti-CMV/TO-Neo-DEST.PLK4 25 N/A pMD2.G VSV-G Addgene #12259 psPAX2 Gag-Pol Addgene #12260 (Continued on next page) ll OPEN ACCESS e2 Current Biology 31, 1403–1416.e1–e7, April 12, 2021 Article .. REAGENT or RESOURCE SOURCE IDENTIFIER Other qEV original Size Exclusion Chromatography (SEC) columns izon #SP1 Tube, Thinwall, Ultra-Clear, 38.5 mL, 25 3 89 mm Beckman coulter #344058 Amicon Ultra-15 Centrifugal Filter Unit Merk #UFC9010 ll OPEN ACCESSArticle

    Bicinchoninic Acid Protein Assay:

    Article Title: Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Alexa-conjugated A488 Life Technologies Cat#A11008; RRID: AB_143165 Mouse Alexa-conjugated A568 Life Technologies Cat#A11001, RRID: AB_2534069 Rabbit Alexa-conjugated A568 Life Technologies Cat#A11011, RRID: AB_143157 Mouse a-tubulin Sigma-Aldrich Cat#T9026; RRID: AB_477593 Rabbit centrin2 N-17-R Santa Cruz Cat#sc-27793-R; RRID: AB_2082359 Mouse LBPA (6C4) Merck Millipore Cat#MABT837 Rabbit LC3B (D11) XP Cell signaling Cat#3868S; RRID: AB_2137707 Mouse a-SMA Sigma-Aldrich Cat#A2547; RRID: AB_476701 Mouse Ki67 Alexa-conjugated A488 BD Biosciences Cat#561165; RRID: AB_10611866 Rabbit TSG101 [(EPR7130(b)] Abcam Cat#ab125011; RRID: AB_10974262 Rabbit CD63 Abcam Cat#ab68418; RRID: AB_10563972 Mouse CD81 (clone B-11) Santa Cruz Biotechnology Cat#sc-166029; RRID: AB_2275892 Mouse ALIX (clone 3A9) Cell signaling Cat#2171, RRID: AB_2299455 Mouse Flotillin-1 (clone 18) Biosciences Cat#610821; RRID: AB_398140 HRP- anti rabbit secondary GE Healthcare Cat#NA934; RRID: AB_772206 HRP- anti mouse secondary GE Healthcare Cat#NA931; RRID: AB_772210 Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma-Aldrich #D9891 H2O2 Sigma-Aldrich #H1009 N-acetyl cysteine Sigma-Aldrich #A9165 Bafilomycin A1 Sigma-Aldrich #B1793-10UG DMEM/F12 Sigma-Aldrich #D8437 DMEM Thermo Fisher Scientific #41966-029 RPMI-1640 Thermo Fisher Scientific #11875093 Keratinocyte serum free medium (1X) Thermo Fisher Scientific #17005042 Opti-MEM reduced serum medium Thermo Fisher Scientific #31985070 DMEM for SILAC Thermo Fisher Scientific #88364 Penicillin/Streptomycin Thermo Fisher Scientific #15140-122 FBS Thermo Fisher Scientific #10500-064 Tet-free FBS Hyclone #SH30070.03T GIBCO FBS, Dialyzed Thermo Fisher Scientific #26400044 Blasticidin Generon #2805-10 Geneticin (G418) Thermo Fisher Scientific #10131027 Puromycin InvivoGen #ant-pr-1 Polybrene Sigma-Aldrich #H9268 Formaldehyde 16% Thermo Fisher Scientific #28908 Alexa Fluor 568 Phalloidin Life Technologies #A12380 Hoechst 33342 ThermoFisher Scientific # H3570 BODIPY FL N-(2-Aminoethyl) Maleimide Thermo Fisher Scientific #B10250 ProLong anti-fade mounting medium Molecular Probes #P36934 BSA Sigma-Aldrich #A9647 Lipofectamine 2000 Invitrogen #11668027 Lipofectamine RNAi Max Invitrogen #13778075 RIPA Buffer Thermo Scientific #89901 Complete Mini Protease Inhibitor Cocktail Roche #11836153001 (Continued on next page) Current Biology 31, 1403–1416.e1–e7, April 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phosphatase inhibitor Cocktail Cell Signaling #5870 Bradford Protein Assay Bio-Rad #5000006 Recombinant Human TGF-b1 PEPROTECH #100-21 Matrigel Matrix Basement Membrane Life Sciences #354234 Corning Rat Tail High Concentration Life Sciences #354249 DTT VWR Chemicals #M109; CAS: 3483-12-3 Iodoacetamide VWR Chemicals Cat#786-228; CAS: 144-48-9 Trypsin Sigma-Aldrich T6567-1MG L-Arginine Sigma-Aldrich A6969-25G L-Lysine Sigma-Aldrich L8662-25G L-Arginine [U-13C6] Cambridge Isotopes CLM-2265-H-0.5 L-Lysine [4,4,5,5-D4] Cambridge Isotopes DLM-2640-0.5 L-Arginine [U-13C6, U-15N4] Cambridge Isotopes CNLM-539-H-0.5 L-Lysine [U-13C6, U-15N2] Cambridge Isotopes CNLM-291-H-0.5 L-Proline Sigma-Aldrich P0380-100G Critical commercial assays GSH/GSSH-Glo assay Promega #V6611 BCA Protein assay Thermo Fisher Scientific #23225 Magic Red Cathepsin B kit Bio-Rad #ICT937 CellVue Maroon Cell Labeling Kit Invitrogen #88-0870-16 Deposited data SILAC analysis of exosome secretion in response to centrosome amplification This paper PRIDE: PXD020984 accessible via PRIDE partner repository (https://www.ebi. ac.uk/pride/archive/) Experimental models: cell lines PaTu-8988T Prof. Hemant Kocher (QMUL) N/A PaTu-8988S Prof. Yaohe Wang (QMUL) N/A Panc-1 Prof. Hemant Kocher (QMUL) N/A CFPAC-1 Prof. David Pellman (Harvard) N/A Capan-1 Prof. Hemant Kocher (QMUL) N/A HPAF-II Prof. Hemant Kocher (QMUL) N/A MIA-PaCa-2 Prof. Hemant Kocher (QMUL) N/A DEC-hTERT Prof. Hemant Kocher (QMUL) N/A HPDE Prof. Yaohe Wang (QMUL) N/A PS1 Prof. Hemant Kocher (QMUL) N/A HEK293M Prof. David Pellman (Harvard) N/A PaTu-8988S.PLK4 This work N/A HPAF-II.PLK4 This work N/A HPAF-II.PLK4 H2B RFP This work N/A PS1 H2B GFP Angus Cameron (QMUL) N/A Oligonucleotides siSAS6 on-TARGET smart pool Dharmacon #M-004158-02 siNegative Control QIAGEN #1027310 Recombinant DNA pLenti-CMV-TetR-Blast Addgene #17492 pLenti-CMV/TO-Neo-DEST.PLK4 25 N/A pMD2.G VSV-G Addgene #12259 psPAX2 Gag-Pol Addgene #12260 (Continued on next page) ll OPEN ACCESS e2 Current Biology 31, 1403–1416.e1–e7, April 12, 2021 Article .. REAGENT or RESOURCE SOURCE IDENTIFIER Other qEV original Size Exclusion Chromatography (SEC) columns izon #SP1 Tube, Thinwall, Ultra-Clear, 38.5 mL, 25 3 89 mm Beckman coulter #344058 Amicon Ultra-15 Centrifugal Filter Unit Merk #UFC9010 ll OPEN ACCESSArticle

    Labeling:

    Article Title: Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Alexa-conjugated A488 Life Technologies Cat#A11008; RRID: AB_143165 Mouse Alexa-conjugated A568 Life Technologies Cat#A11001, RRID: AB_2534069 Rabbit Alexa-conjugated A568 Life Technologies Cat#A11011, RRID: AB_143157 Mouse a-tubulin Sigma-Aldrich Cat#T9026; RRID: AB_477593 Rabbit centrin2 N-17-R Santa Cruz Cat#sc-27793-R; RRID: AB_2082359 Mouse LBPA (6C4) Merck Millipore Cat#MABT837 Rabbit LC3B (D11) XP Cell signaling Cat#3868S; RRID: AB_2137707 Mouse a-SMA Sigma-Aldrich Cat#A2547; RRID: AB_476701 Mouse Ki67 Alexa-conjugated A488 BD Biosciences Cat#561165; RRID: AB_10611866 Rabbit TSG101 [(EPR7130(b)] Abcam Cat#ab125011; RRID: AB_10974262 Rabbit CD63 Abcam Cat#ab68418; RRID: AB_10563972 Mouse CD81 (clone B-11) Santa Cruz Biotechnology Cat#sc-166029; RRID: AB_2275892 Mouse ALIX (clone 3A9) Cell signaling Cat#2171, RRID: AB_2299455 Mouse Flotillin-1 (clone 18) Biosciences Cat#610821; RRID: AB_398140 HRP- anti rabbit secondary GE Healthcare Cat#NA934; RRID: AB_772206 HRP- anti mouse secondary GE Healthcare Cat#NA931; RRID: AB_772210 Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma-Aldrich #D9891 H2O2 Sigma-Aldrich #H1009 N-acetyl cysteine Sigma-Aldrich #A9165 Bafilomycin A1 Sigma-Aldrich #B1793-10UG DMEM/F12 Sigma-Aldrich #D8437 DMEM Thermo Fisher Scientific #41966-029 RPMI-1640 Thermo Fisher Scientific #11875093 Keratinocyte serum free medium (1X) Thermo Fisher Scientific #17005042 Opti-MEM reduced serum medium Thermo Fisher Scientific #31985070 DMEM for SILAC Thermo Fisher Scientific #88364 Penicillin/Streptomycin Thermo Fisher Scientific #15140-122 FBS Thermo Fisher Scientific #10500-064 Tet-free FBS Hyclone #SH30070.03T GIBCO FBS, Dialyzed Thermo Fisher Scientific #26400044 Blasticidin Generon #2805-10 Geneticin (G418) Thermo Fisher Scientific #10131027 Puromycin InvivoGen #ant-pr-1 Polybrene Sigma-Aldrich #H9268 Formaldehyde 16% Thermo Fisher Scientific #28908 Alexa Fluor 568 Phalloidin Life Technologies #A12380 Hoechst 33342 ThermoFisher Scientific # H3570 BODIPY FL N-(2-Aminoethyl) Maleimide Thermo Fisher Scientific #B10250 ProLong anti-fade mounting medium Molecular Probes #P36934 BSA Sigma-Aldrich #A9647 Lipofectamine 2000 Invitrogen #11668027 Lipofectamine RNAi Max Invitrogen #13778075 RIPA Buffer Thermo Scientific #89901 Complete Mini Protease Inhibitor Cocktail Roche #11836153001 (Continued on next page) Current Biology 31, 1403–1416.e1–e7, April 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phosphatase inhibitor Cocktail Cell Signaling #5870 Bradford Protein Assay Bio-Rad #5000006 Recombinant Human TGF-b1 PEPROTECH #100-21 Matrigel Matrix Basement Membrane Life Sciences #354234 Corning Rat Tail High Concentration Life Sciences #354249 DTT VWR Chemicals #M109; CAS: 3483-12-3 Iodoacetamide VWR Chemicals Cat#786-228; CAS: 144-48-9 Trypsin Sigma-Aldrich T6567-1MG L-Arginine Sigma-Aldrich A6969-25G L-Lysine Sigma-Aldrich L8662-25G L-Arginine [U-13C6] Cambridge Isotopes CLM-2265-H-0.5 L-Lysine [4,4,5,5-D4] Cambridge Isotopes DLM-2640-0.5 L-Arginine [U-13C6, U-15N4] Cambridge Isotopes CNLM-539-H-0.5 L-Lysine [U-13C6, U-15N2] Cambridge Isotopes CNLM-291-H-0.5 L-Proline Sigma-Aldrich P0380-100G Critical commercial assays GSH/GSSH-Glo assay Promega #V6611 BCA Protein assay Thermo Fisher Scientific #23225 Magic Red Cathepsin B kit Bio-Rad #ICT937 CellVue Maroon Cell Labeling Kit Invitrogen #88-0870-16 Deposited data SILAC analysis of exosome secretion in response to centrosome amplification This paper PRIDE: PXD020984 accessible via PRIDE partner repository (https://www.ebi. ac.uk/pride/archive/) Experimental models: cell lines PaTu-8988T Prof. Hemant Kocher (QMUL) N/A PaTu-8988S Prof. Yaohe Wang (QMUL) N/A Panc-1 Prof. Hemant Kocher (QMUL) N/A CFPAC-1 Prof. David Pellman (Harvard) N/A Capan-1 Prof. Hemant Kocher (QMUL) N/A HPAF-II Prof. Hemant Kocher (QMUL) N/A MIA-PaCa-2 Prof. Hemant Kocher (QMUL) N/A DEC-hTERT Prof. Hemant Kocher (QMUL) N/A HPDE Prof. Yaohe Wang (QMUL) N/A PS1 Prof. Hemant Kocher (QMUL) N/A HEK293M Prof. David Pellman (Harvard) N/A PaTu-8988S.PLK4 This work N/A HPAF-II.PLK4 This work N/A HPAF-II.PLK4 H2B RFP This work N/A PS1 H2B GFP Angus Cameron (QMUL) N/A Oligonucleotides siSAS6 on-TARGET smart pool Dharmacon #M-004158-02 siNegative Control QIAGEN #1027310 Recombinant DNA pLenti-CMV-TetR-Blast Addgene #17492 pLenti-CMV/TO-Neo-DEST.PLK4 25 N/A pMD2.G VSV-G Addgene #12259 psPAX2 Gag-Pol Addgene #12260 (Continued on next page) ll OPEN ACCESS e2 Current Biology 31, 1403–1416.e1–e7, April 12, 2021 Article .. REAGENT or RESOURCE SOURCE IDENTIFIER Other qEV original Size Exclusion Chromatography (SEC) columns izon #SP1 Tube, Thinwall, Ultra-Clear, 38.5 mL, 25 3 89 mm Beckman coulter #344058 Amicon Ultra-15 Centrifugal Filter Unit Merk #UFC9010 ll OPEN ACCESSArticle

    Multiplex sample analysis:

    Article Title: Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Alexa-conjugated A488 Life Technologies Cat#A11008; RRID: AB_143165 Mouse Alexa-conjugated A568 Life Technologies Cat#A11001, RRID: AB_2534069 Rabbit Alexa-conjugated A568 Life Technologies Cat#A11011, RRID: AB_143157 Mouse a-tubulin Sigma-Aldrich Cat#T9026; RRID: AB_477593 Rabbit centrin2 N-17-R Santa Cruz Cat#sc-27793-R; RRID: AB_2082359 Mouse LBPA (6C4) Merck Millipore Cat#MABT837 Rabbit LC3B (D11) XP Cell signaling Cat#3868S; RRID: AB_2137707 Mouse a-SMA Sigma-Aldrich Cat#A2547; RRID: AB_476701 Mouse Ki67 Alexa-conjugated A488 BD Biosciences Cat#561165; RRID: AB_10611866 Rabbit TSG101 [(EPR7130(b)] Abcam Cat#ab125011; RRID: AB_10974262 Rabbit CD63 Abcam Cat#ab68418; RRID: AB_10563972 Mouse CD81 (clone B-11) Santa Cruz Biotechnology Cat#sc-166029; RRID: AB_2275892 Mouse ALIX (clone 3A9) Cell signaling Cat#2171, RRID: AB_2299455 Mouse Flotillin-1 (clone 18) Biosciences Cat#610821; RRID: AB_398140 HRP- anti rabbit secondary GE Healthcare Cat#NA934; RRID: AB_772206 HRP- anti mouse secondary GE Healthcare Cat#NA931; RRID: AB_772210 Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma-Aldrich #D9891 H2O2 Sigma-Aldrich #H1009 N-acetyl cysteine Sigma-Aldrich #A9165 Bafilomycin A1 Sigma-Aldrich #B1793-10UG DMEM/F12 Sigma-Aldrich #D8437 DMEM Thermo Fisher Scientific #41966-029 RPMI-1640 Thermo Fisher Scientific #11875093 Keratinocyte serum free medium (1X) Thermo Fisher Scientific #17005042 Opti-MEM reduced serum medium Thermo Fisher Scientific #31985070 DMEM for SILAC Thermo Fisher Scientific #88364 Penicillin/Streptomycin Thermo Fisher Scientific #15140-122 FBS Thermo Fisher Scientific #10500-064 Tet-free FBS Hyclone #SH30070.03T GIBCO FBS, Dialyzed Thermo Fisher Scientific #26400044 Blasticidin Generon #2805-10 Geneticin (G418) Thermo Fisher Scientific #10131027 Puromycin InvivoGen #ant-pr-1 Polybrene Sigma-Aldrich #H9268 Formaldehyde 16% Thermo Fisher Scientific #28908 Alexa Fluor 568 Phalloidin Life Technologies #A12380 Hoechst 33342 ThermoFisher Scientific # H3570 BODIPY FL N-(2-Aminoethyl) Maleimide Thermo Fisher Scientific #B10250 ProLong anti-fade mounting medium Molecular Probes #P36934 BSA Sigma-Aldrich #A9647 Lipofectamine 2000 Invitrogen #11668027 Lipofectamine RNAi Max Invitrogen #13778075 RIPA Buffer Thermo Scientific #89901 Complete Mini Protease Inhibitor Cocktail Roche #11836153001 (Continued on next page) Current Biology 31, 1403–1416.e1–e7, April 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phosphatase inhibitor Cocktail Cell Signaling #5870 Bradford Protein Assay Bio-Rad #5000006 Recombinant Human TGF-b1 PEPROTECH #100-21 Matrigel Matrix Basement Membrane Life Sciences #354234 Corning Rat Tail High Concentration Life Sciences #354249 DTT VWR Chemicals #M109; CAS: 3483-12-3 Iodoacetamide VWR Chemicals Cat#786-228; CAS: 144-48-9 Trypsin Sigma-Aldrich T6567-1MG L-Arginine Sigma-Aldrich A6969-25G L-Lysine Sigma-Aldrich L8662-25G L-Arginine [U-13C6] Cambridge Isotopes CLM-2265-H-0.5 L-Lysine [4,4,5,5-D4] Cambridge Isotopes DLM-2640-0.5 L-Arginine [U-13C6, U-15N4] Cambridge Isotopes CNLM-539-H-0.5 L-Lysine [U-13C6, U-15N2] Cambridge Isotopes CNLM-291-H-0.5 L-Proline Sigma-Aldrich P0380-100G Critical commercial assays GSH/GSSH-Glo assay Promega #V6611 BCA Protein assay Thermo Fisher Scientific #23225 Magic Red Cathepsin B kit Bio-Rad #ICT937 CellVue Maroon Cell Labeling Kit Invitrogen #88-0870-16 Deposited data SILAC analysis of exosome secretion in response to centrosome amplification This paper PRIDE: PXD020984 accessible via PRIDE partner repository (https://www.ebi. ac.uk/pride/archive/) Experimental models: cell lines PaTu-8988T Prof. Hemant Kocher (QMUL) N/A PaTu-8988S Prof. Yaohe Wang (QMUL) N/A Panc-1 Prof. Hemant Kocher (QMUL) N/A CFPAC-1 Prof. David Pellman (Harvard) N/A Capan-1 Prof. Hemant Kocher (QMUL) N/A HPAF-II Prof. Hemant Kocher (QMUL) N/A MIA-PaCa-2 Prof. Hemant Kocher (QMUL) N/A DEC-hTERT Prof. Hemant Kocher (QMUL) N/A HPDE Prof. Yaohe Wang (QMUL) N/A PS1 Prof. Hemant Kocher (QMUL) N/A HEK293M Prof. David Pellman (Harvard) N/A PaTu-8988S.PLK4 This work N/A HPAF-II.PLK4 This work N/A HPAF-II.PLK4 H2B RFP This work N/A PS1 H2B GFP Angus Cameron (QMUL) N/A Oligonucleotides siSAS6 on-TARGET smart pool Dharmacon #M-004158-02 siNegative Control QIAGEN #1027310 Recombinant DNA pLenti-CMV-TetR-Blast Addgene #17492 pLenti-CMV/TO-Neo-DEST.PLK4 25 N/A pMD2.G VSV-G Addgene #12259 psPAX2 Gag-Pol Addgene #12260 (Continued on next page) ll OPEN ACCESS e2 Current Biology 31, 1403–1416.e1–e7, April 12, 2021 Article .. REAGENT or RESOURCE SOURCE IDENTIFIER Other qEV original Size Exclusion Chromatography (SEC) columns izon #SP1 Tube, Thinwall, Ultra-Clear, 38.5 mL, 25 3 89 mm Beckman coulter #344058 Amicon Ultra-15 Centrifugal Filter Unit Merk #UFC9010 ll OPEN ACCESSArticle

    Amplification:

    Article Title: Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Alexa-conjugated A488 Life Technologies Cat#A11008; RRID: AB_143165 Mouse Alexa-conjugated A568 Life Technologies Cat#A11001, RRID: AB_2534069 Rabbit Alexa-conjugated A568 Life Technologies Cat#A11011, RRID: AB_143157 Mouse a-tubulin Sigma-Aldrich Cat#T9026; RRID: AB_477593 Rabbit centrin2 N-17-R Santa Cruz Cat#sc-27793-R; RRID: AB_2082359 Mouse LBPA (6C4) Merck Millipore Cat#MABT837 Rabbit LC3B (D11) XP Cell signaling Cat#3868S; RRID: AB_2137707 Mouse a-SMA Sigma-Aldrich Cat#A2547; RRID: AB_476701 Mouse Ki67 Alexa-conjugated A488 BD Biosciences Cat#561165; RRID: AB_10611866 Rabbit TSG101 [(EPR7130(b)] Abcam Cat#ab125011; RRID: AB_10974262 Rabbit CD63 Abcam Cat#ab68418; RRID: AB_10563972 Mouse CD81 (clone B-11) Santa Cruz Biotechnology Cat#sc-166029; RRID: AB_2275892 Mouse ALIX (clone 3A9) Cell signaling Cat#2171, RRID: AB_2299455 Mouse Flotillin-1 (clone 18) Biosciences Cat#610821; RRID: AB_398140 HRP- anti rabbit secondary GE Healthcare Cat#NA934; RRID: AB_772206 HRP- anti mouse secondary GE Healthcare Cat#NA931; RRID: AB_772210 Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma-Aldrich #D9891 H2O2 Sigma-Aldrich #H1009 N-acetyl cysteine Sigma-Aldrich #A9165 Bafilomycin A1 Sigma-Aldrich #B1793-10UG DMEM/F12 Sigma-Aldrich #D8437 DMEM Thermo Fisher Scientific #41966-029 RPMI-1640 Thermo Fisher Scientific #11875093 Keratinocyte serum free medium (1X) Thermo Fisher Scientific #17005042 Opti-MEM reduced serum medium Thermo Fisher Scientific #31985070 DMEM for SILAC Thermo Fisher Scientific #88364 Penicillin/Streptomycin Thermo Fisher Scientific #15140-122 FBS Thermo Fisher Scientific #10500-064 Tet-free FBS Hyclone #SH30070.03T GIBCO FBS, Dialyzed Thermo Fisher Scientific #26400044 Blasticidin Generon #2805-10 Geneticin (G418) Thermo Fisher Scientific #10131027 Puromycin InvivoGen #ant-pr-1 Polybrene Sigma-Aldrich #H9268 Formaldehyde 16% Thermo Fisher Scientific #28908 Alexa Fluor 568 Phalloidin Life Technologies #A12380 Hoechst 33342 ThermoFisher Scientific # H3570 BODIPY FL N-(2-Aminoethyl) Maleimide Thermo Fisher Scientific #B10250 ProLong anti-fade mounting medium Molecular Probes #P36934 BSA Sigma-Aldrich #A9647 Lipofectamine 2000 Invitrogen #11668027 Lipofectamine RNAi Max Invitrogen #13778075 RIPA Buffer Thermo Scientific #89901 Complete Mini Protease Inhibitor Cocktail Roche #11836153001 (Continued on next page) Current Biology 31, 1403–1416.e1–e7, April 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phosphatase inhibitor Cocktail Cell Signaling #5870 Bradford Protein Assay Bio-Rad #5000006 Recombinant Human TGF-b1 PEPROTECH #100-21 Matrigel Matrix Basement Membrane Life Sciences #354234 Corning Rat Tail High Concentration Life Sciences #354249 DTT VWR Chemicals #M109; CAS: 3483-12-3 Iodoacetamide VWR Chemicals Cat#786-228; CAS: 144-48-9 Trypsin Sigma-Aldrich T6567-1MG L-Arginine Sigma-Aldrich A6969-25G L-Lysine Sigma-Aldrich L8662-25G L-Arginine [U-13C6] Cambridge Isotopes CLM-2265-H-0.5 L-Lysine [4,4,5,5-D4] Cambridge Isotopes DLM-2640-0.5 L-Arginine [U-13C6, U-15N4] Cambridge Isotopes CNLM-539-H-0.5 L-Lysine [U-13C6, U-15N2] Cambridge Isotopes CNLM-291-H-0.5 L-Proline Sigma-Aldrich P0380-100G Critical commercial assays GSH/GSSH-Glo assay Promega #V6611 BCA Protein assay Thermo Fisher Scientific #23225 Magic Red Cathepsin B kit Bio-Rad #ICT937 CellVue Maroon Cell Labeling Kit Invitrogen #88-0870-16 Deposited data SILAC analysis of exosome secretion in response to centrosome amplification This paper PRIDE: PXD020984 accessible via PRIDE partner repository (https://www.ebi. ac.uk/pride/archive/) Experimental models: cell lines PaTu-8988T Prof. Hemant Kocher (QMUL) N/A PaTu-8988S Prof. Yaohe Wang (QMUL) N/A Panc-1 Prof. Hemant Kocher (QMUL) N/A CFPAC-1 Prof. David Pellman (Harvard) N/A Capan-1 Prof. Hemant Kocher (QMUL) N/A HPAF-II Prof. Hemant Kocher (QMUL) N/A MIA-PaCa-2 Prof. Hemant Kocher (QMUL) N/A DEC-hTERT Prof. Hemant Kocher (QMUL) N/A HPDE Prof. Yaohe Wang (QMUL) N/A PS1 Prof. Hemant Kocher (QMUL) N/A HEK293M Prof. David Pellman (Harvard) N/A PaTu-8988S.PLK4 This work N/A HPAF-II.PLK4 This work N/A HPAF-II.PLK4 H2B RFP This work N/A PS1 H2B GFP Angus Cameron (QMUL) N/A Oligonucleotides siSAS6 on-TARGET smart pool Dharmacon #M-004158-02 siNegative Control QIAGEN #1027310 Recombinant DNA pLenti-CMV-TetR-Blast Addgene #17492 pLenti-CMV/TO-Neo-DEST.PLK4 25 N/A pMD2.G VSV-G Addgene #12259 psPAX2 Gag-Pol Addgene #12260 (Continued on next page) ll OPEN ACCESS e2 Current Biology 31, 1403–1416.e1–e7, April 12, 2021 Article .. REAGENT or RESOURCE SOURCE IDENTIFIER Other qEV original Size Exclusion Chromatography (SEC) columns izon #SP1 Tube, Thinwall, Ultra-Clear, 38.5 mL, 25 3 89 mm Beckman coulter #344058 Amicon Ultra-15 Centrifugal Filter Unit Merk #UFC9010 ll OPEN ACCESSArticle

    Control:

    Article Title: Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Alexa-conjugated A488 Life Technologies Cat#A11008; RRID: AB_143165 Mouse Alexa-conjugated A568 Life Technologies Cat#A11001, RRID: AB_2534069 Rabbit Alexa-conjugated A568 Life Technologies Cat#A11011, RRID: AB_143157 Mouse a-tubulin Sigma-Aldrich Cat#T9026; RRID: AB_477593 Rabbit centrin2 N-17-R Santa Cruz Cat#sc-27793-R; RRID: AB_2082359 Mouse LBPA (6C4) Merck Millipore Cat#MABT837 Rabbit LC3B (D11) XP Cell signaling Cat#3868S; RRID: AB_2137707 Mouse a-SMA Sigma-Aldrich Cat#A2547; RRID: AB_476701 Mouse Ki67 Alexa-conjugated A488 BD Biosciences Cat#561165; RRID: AB_10611866 Rabbit TSG101 [(EPR7130(b)] Abcam Cat#ab125011; RRID: AB_10974262 Rabbit CD63 Abcam Cat#ab68418; RRID: AB_10563972 Mouse CD81 (clone B-11) Santa Cruz Biotechnology Cat#sc-166029; RRID: AB_2275892 Mouse ALIX (clone 3A9) Cell signaling Cat#2171, RRID: AB_2299455 Mouse Flotillin-1 (clone 18) Biosciences Cat#610821; RRID: AB_398140 HRP- anti rabbit secondary GE Healthcare Cat#NA934; RRID: AB_772206 HRP- anti mouse secondary GE Healthcare Cat#NA931; RRID: AB_772210 Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma-Aldrich #D9891 H2O2 Sigma-Aldrich #H1009 N-acetyl cysteine Sigma-Aldrich #A9165 Bafilomycin A1 Sigma-Aldrich #B1793-10UG DMEM/F12 Sigma-Aldrich #D8437 DMEM Thermo Fisher Scientific #41966-029 RPMI-1640 Thermo Fisher Scientific #11875093 Keratinocyte serum free medium (1X) Thermo Fisher Scientific #17005042 Opti-MEM reduced serum medium Thermo Fisher Scientific #31985070 DMEM for SILAC Thermo Fisher Scientific #88364 Penicillin/Streptomycin Thermo Fisher Scientific #15140-122 FBS Thermo Fisher Scientific #10500-064 Tet-free FBS Hyclone #SH30070.03T GIBCO FBS, Dialyzed Thermo Fisher Scientific #26400044 Blasticidin Generon #2805-10 Geneticin (G418) Thermo Fisher Scientific #10131027 Puromycin InvivoGen #ant-pr-1 Polybrene Sigma-Aldrich #H9268 Formaldehyde 16% Thermo Fisher Scientific #28908 Alexa Fluor 568 Phalloidin Life Technologies #A12380 Hoechst 33342 ThermoFisher Scientific # H3570 BODIPY FL N-(2-Aminoethyl) Maleimide Thermo Fisher Scientific #B10250 ProLong anti-fade mounting medium Molecular Probes #P36934 BSA Sigma-Aldrich #A9647 Lipofectamine 2000 Invitrogen #11668027 Lipofectamine RNAi Max Invitrogen #13778075 RIPA Buffer Thermo Scientific #89901 Complete Mini Protease Inhibitor Cocktail Roche #11836153001 (Continued on next page) Current Biology 31, 1403–1416.e1–e7, April 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phosphatase inhibitor Cocktail Cell Signaling #5870 Bradford Protein Assay Bio-Rad #5000006 Recombinant Human TGF-b1 PEPROTECH #100-21 Matrigel Matrix Basement Membrane Life Sciences #354234 Corning Rat Tail High Concentration Life Sciences #354249 DTT VWR Chemicals #M109; CAS: 3483-12-3 Iodoacetamide VWR Chemicals Cat#786-228; CAS: 144-48-9 Trypsin Sigma-Aldrich T6567-1MG L-Arginine Sigma-Aldrich A6969-25G L-Lysine Sigma-Aldrich L8662-25G L-Arginine [U-13C6] Cambridge Isotopes CLM-2265-H-0.5 L-Lysine [4,4,5,5-D4] Cambridge Isotopes DLM-2640-0.5 L-Arginine [U-13C6, U-15N4] Cambridge Isotopes CNLM-539-H-0.5 L-Lysine [U-13C6, U-15N2] Cambridge Isotopes CNLM-291-H-0.5 L-Proline Sigma-Aldrich P0380-100G Critical commercial assays GSH/GSSH-Glo assay Promega #V6611 BCA Protein assay Thermo Fisher Scientific #23225 Magic Red Cathepsin B kit Bio-Rad #ICT937 CellVue Maroon Cell Labeling Kit Invitrogen #88-0870-16 Deposited data SILAC analysis of exosome secretion in response to centrosome amplification This paper PRIDE: PXD020984 accessible via PRIDE partner repository (https://www.ebi. ac.uk/pride/archive/) Experimental models: cell lines PaTu-8988T Prof. Hemant Kocher (QMUL) N/A PaTu-8988S Prof. Yaohe Wang (QMUL) N/A Panc-1 Prof. Hemant Kocher (QMUL) N/A CFPAC-1 Prof. David Pellman (Harvard) N/A Capan-1 Prof. Hemant Kocher (QMUL) N/A HPAF-II Prof. Hemant Kocher (QMUL) N/A MIA-PaCa-2 Prof. Hemant Kocher (QMUL) N/A DEC-hTERT Prof. Hemant Kocher (QMUL) N/A HPDE Prof. Yaohe Wang (QMUL) N/A PS1 Prof. Hemant Kocher (QMUL) N/A HEK293M Prof. David Pellman (Harvard) N/A PaTu-8988S.PLK4 This work N/A HPAF-II.PLK4 This work N/A HPAF-II.PLK4 H2B RFP This work N/A PS1 H2B GFP Angus Cameron (QMUL) N/A Oligonucleotides siSAS6 on-TARGET smart pool Dharmacon #M-004158-02 siNegative Control QIAGEN #1027310 Recombinant DNA pLenti-CMV-TetR-Blast Addgene #17492 pLenti-CMV/TO-Neo-DEST.PLK4 25 N/A pMD2.G VSV-G Addgene #12259 psPAX2 Gag-Pol Addgene #12260 (Continued on next page) ll OPEN ACCESS e2 Current Biology 31, 1403–1416.e1–e7, April 12, 2021 Article .. REAGENT or RESOURCE SOURCE IDENTIFIER Other qEV original Size Exclusion Chromatography (SEC) columns izon #SP1 Tube, Thinwall, Ultra-Clear, 38.5 mL, 25 3 89 mm Beckman coulter #344058 Amicon Ultra-15 Centrifugal Filter Unit Merk #UFC9010 ll OPEN ACCESSArticle

    Sequencing:

    Article Title: PARP inhibitors suppress tumours via centrosome error-induced senescence independent of DNA damage response
    Article Snippet: .. For inducible PLK4 overexpression in MCF7 cells, the human PLK4 coding sequence was subcloned into pLenti-CMV/TO-Neo-Dest (Addgene, Cat# 17292). .. For Flag-BRCA1 or Flag-ZNF721 overexpression in MCF7 cells, the human BRCA1 or ZNF721 coding sequence was subcloned into pcDNA3.1-Flag (Addgene, Cat# 208051).

    Article Title: PARP inhibitors suppress tumours via centrosome error-induced senescence independent of DNA damage response.
    Article Snippet: .. For inducible PLK4 overexpression in MCF7 cells, the human PLK4 coding sequence was subcloned into pLenti-CMV/TO-Neo-Dest (Addgene, Cat# 17292). .. For Flag-BRCA1 or Flag-ZNF721 overexpression in MCF7 cells, the human BRCA1 or ZNF721 coding sequence was subcloned into pcDNA3.1-Flag (Addgene, Cat# 208051).



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    (A) Schematic of injection for labelling of RSC→V1 axons and two-photon recording. (B) Example field of view showing RSC axons in layer 1 of V1 labelled with jGCaMP8s. (C) Example spatial responses (average traces plotted over heatmap) for three visually responsive boutons. (D) 2d Gaussian fits of the spatial receptive fields of boutons shown in C. (E) Distribution of spatial receptive field size (sigma from fit) in azimuth and elevation. (F) Distribution of fitted preferred position of each RSC→V1 bouton relative to the preferred position of other boutons in the same field of view. n = 1950 boutons collected from 8 mice in 12 experiments.

    Journal: bioRxiv

    Article Title: The functional organisation of retrosplenial feedback to V1

    doi: 10.1101/2025.09.25.678583

    Figure Lengend Snippet: (A) Schematic of injection for labelling of RSC→V1 axons and two-photon recording. (B) Example field of view showing RSC axons in layer 1 of V1 labelled with jGCaMP8s. (C) Example spatial responses (average traces plotted over heatmap) for three visually responsive boutons. (D) 2d Gaussian fits of the spatial receptive fields of boutons shown in C. (E) Distribution of spatial receptive field size (sigma from fit) in azimuth and elevation. (F) Distribution of fitted preferred position of each RSC→V1 bouton relative to the preferred position of other boutons in the same field of view. n = 1950 boutons collected from 8 mice in 12 experiments.

    Article Snippet: After the brain was exposed, an AAV was injected at a depth of 400μm to drive expression of jGCaMP8s (pAAV-hSynapsin1-axon-jGCaMP8s-P2A-mRuby3, Addgene plasmid #172921, 5 × 10 12 GC per millilitre titre, volume 100 nL).

    Techniques: Injection

    (A) Schematic of injection for labelling of V1 somas with jRGECO1a and RSC→V1 axons with jGCaMP8s and two-photon recording. (B) Example field of view showing RSC axons in layer 1 of V1 labelled with jGCaMP8s and layer 2/3 somas labelled with jRGECO1a. (C) Example spatial responses (average traces plotted over heatmap) for three visually responsive boutons (left) and 3 retinotopically aligned V1 somas (right). (D) Heat plot showing distribution of difference in tuning of all RSC→V1 boutons relative to local V1 neurons in azimuth and elevation. (E) Responses of each RSC→V1 bouton to expanding versus contracting stimulus. (F) Data from D divided into binocular and monocular areas of the V1 retinotopic map showing nasal offset of RSC→V1 bouton positional tuning relative to V1 tuning in monocular area. (G) Distribution of ΔV1-RSC bouton positional preference showing offset is specific to monocular visual space. For V1 soma recordings, n = 827 neurons from 9 experiments in 6 mice. For RSC→V1 bouton recordings, n = 1467 boutons from 9 experiments in the same 6 mice used for soma recordings.

    Journal: bioRxiv

    Article Title: The functional organisation of retrosplenial feedback to V1

    doi: 10.1101/2025.09.25.678583

    Figure Lengend Snippet: (A) Schematic of injection for labelling of V1 somas with jRGECO1a and RSC→V1 axons with jGCaMP8s and two-photon recording. (B) Example field of view showing RSC axons in layer 1 of V1 labelled with jGCaMP8s and layer 2/3 somas labelled with jRGECO1a. (C) Example spatial responses (average traces plotted over heatmap) for three visually responsive boutons (left) and 3 retinotopically aligned V1 somas (right). (D) Heat plot showing distribution of difference in tuning of all RSC→V1 boutons relative to local V1 neurons in azimuth and elevation. (E) Responses of each RSC→V1 bouton to expanding versus contracting stimulus. (F) Data from D divided into binocular and monocular areas of the V1 retinotopic map showing nasal offset of RSC→V1 bouton positional tuning relative to V1 tuning in monocular area. (G) Distribution of ΔV1-RSC bouton positional preference showing offset is specific to monocular visual space. For V1 soma recordings, n = 827 neurons from 9 experiments in 6 mice. For RSC→V1 bouton recordings, n = 1467 boutons from 9 experiments in the same 6 mice used for soma recordings.

    Article Snippet: After the brain was exposed, an AAV was injected at a depth of 400μm to drive expression of jGCaMP8s (pAAV-hSynapsin1-axon-jGCaMP8s-P2A-mRuby3, Addgene plasmid #172921, 5 × 10 12 GC per millilitre titre, volume 100 nL).

    Techniques: Injection