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  • br based approach to model patterns of

    2019-10-30


    based approach to model patterns Aztreonam chemoresistance and tumor recurrence using ovarian cancer stem cell spheroids. Clin. Cancer Res. 23, 6934–6945.
    Weerapana, E., Speers, A.E., and Cravatt, B.F. (2007). Tandem orthogonal proteolysis-activity-based protein profiling (TOP-ABPP)—a general method for mapping sites of probe modification in proteomes. Nat. Protoc 2, 1414– 1425.
    STAR+METHODS
    KEY RESOURCES TABLE
    REAGENT or RESOURCE
    SOURCE IDENTIFIER
    Antibodies
    Anti ALDH1A1
    Novus
    Anti ALDH1A3
    Novus
    Anti UCP1
    Anti UCP3
    Abcam
    Anti RIPK1
    Cell Signaling
    Anti RIPK3
    Cell Signaling
    Anti PGAM5
    GeneTex
    Anti p-DRP1
    GeneTex
    Anti DRP1
    Cell Signaling
    Anti b-tubulin
    Cell Signaling
    Anti TOM20
    Cell Signaling
    Anti ACTB
    Cell Signaling
    Anti GAPDH
    Cell Signaling
    Anti Na1K-ATPase1
    Cell Signaling
    Anti FADD
    Sigma-Aldrich
    Miltenyl Biotec
    Bacterial and Virus Strains
    Subcloning Efficiency DH5a Competent Cells
    Invitrogen
    Biological Samples
    Patient-derived xenografts PDXs
    This paper
    Charles Landen lab
    Patient tumor samples
    This paper
    Buckanovich lab
    Chemicals, Peptides, and Recombinant Proteins
    Pan-Caspase inhibitor Z-VAD-FMK
    InvivoGen
    tlrl-vad
    Caspase 3 Inhibitor Z-DEVD-FMK
    BAPTA
    Tocris
    Necrostatin-1
    Tocris
    Chembridge
    Hurley Lab
    Necrosulfonamide
    Tocris
    Critical Commercial Assays
    Aldefluor kit
    Stem cell
    Fluo Calcium Indicator CM-H2DCF
    Invitrogen
    SuperScript III First strand synthesis SuperMix
    Invitrogen
    Deposited Data
    CCLE cell line analysis-ALDH1A expression
    This paper
    Table S1
    673A PubChem search
    This paper
    Table S2
    CM10 PubChem search
    This paper
    Table S3
    Experimental Models: Cell Lines
    Ovsaho, Kuramochi
    JCRB bank
    JCRB bank
    ATCC
    Duke
    Experimental Models: Organisms/Strains
    NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ
    The Jackson labs
    LSL-K-rasG12D/+PtenloxP/loxP
    Romero lab
    Romero lab
    (Continued on next page)
    Continued
    REAGENT or RESOURCE
    SOURCE IDENTIFIER
    Oligonucleotides
    siALDH1A1
    siALDH1A2
    siALDH1A3
    Sigma
    siUCP1
    Dharmacon
    siUCP3
    siRIPK1
    Dharmacon
    siRIPK3
    ABM
    Recombinant DNA
    UCP1 plasmid
    TransOmics clone BC008392
    UCP3 plasmid
    TransOmics Clone BC096
    CONTACT FOR REAGENT AND RESOURCE SHARING
    Further information and requests for resources and reagents should be directed to and will be fulfilled by the Lead Contact, Ron Buckanovich ([email protected]).
    EXPERIMENTAL MODEL AND SUBJECT DETAIL
    2) Primary samples: primary ovarian cancer tumors were obtained after surgery and dissociated to single cell suspension. Single cell suspension was used for spheroid studies.
    3) LSL-K-ras G12D/+ Pten loxP/loxP was established by Dr. Romero, OBGYN department, University of Chicago
    4) PDX model of ovarian cancer was established by Dr. Charles Landen, OBGYN department, University of Virginia and Dr. Buck-anovich University of Michigan.
    5) NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ mice from Jackson labs #00557 were used for all subcutaneous and IP injections
    METHOD DETAILS
    Cell culture and reagents
    PEO1, PEO4, OvCar8, and Kuramochi were used in experiments that assessed 673A effect on CD133 cells due to very high CD133+ expression. A2780, Ovsaho, and OVCAR5 cells were used in experiments which required very distinctive CD133+ population. SKoV3 were used in experiments that required very bright ALDH+ population. OvCar5 and OvCar8 used due to their high ALDH1A3 expression.