# Test file for EGFRSignalingAssay
egfr_signaling_assays:
  - id: "EGFR:001"
    name: "EGFR phosphorylation after smoke exposure"
    has_specified_output:
      id: "EGFR:001-output"
      name: "EGFR:001 measurement results"
      egfr_phosphorylation_y1068:
        value: "3.2"
        unit:
          id: "UO:0000193"
          name: "fold change"
      total_egfr_protein:
        value: "1.0"
        unit:
          id: "UO:0000193"
          name: "fold change"
      egfr_phosphorylation_y1173:
        value: "2.1"
        unit:
          id: "UO:0000193"
          name: "fold change"
      downstream_kinase_activation: "ERK1/2 and AKT activated"
      erk_phosphorylation:
        value: "2.8"
        unit:
          id: "UO:0000193"
          name: "fold change"
      akt_phosphorylation:
        value: "1.5"
        unit:
          id: "UO:0000193"
          name: "fold change"
      egfr_ligand_expression:
        value: "1.9"
        unit:
          id: "UO:0000193"
          name: "fold change"
      egfr_membrane_localization:
        value: "78"
        unit:
          id: "UO:0000187"
          name: "percent"
    normalization_reference: "beta-actin"
    assay_date: "2024-05-08"
    informs_on_key_event:
      id: "KE:ke1-egfr-activation"
      name: "EGFR activation"
      biological_action: activated
      level_of_biological_organization: molecular
    has_exposure_condition:
      - id: "EXPOSURE:cse-4h"
        name: "Cigarette smoke extract 5% for 4 hours"
        exposure_agent:
          id: "CHEBI:39188"
          name: "cigarette smoke extract"
        exposure_concentration:
          value: "5"
          unit:
            id: "UO:0000187"
            name: "percent"
        exposure_duration:
          value: "4"
          unit:
            id: "UO:0000032"
            name: "hour"
    study_subject:
      subject_type: CellularSystem
      id: "soma:culture-005"
      name: "Primary HBE ALI culture"
      cell_type:
        id: "CL:0002328"
        name: "bronchial epithelial cell"
      model_species:
        id: "NCBITaxon:9606"
        name: "Homo sapiens"
    follows_protocols:
      - protocol_type: MolecularAssayProtocol
        id: "PROTOCOL:wb-001"
        name: "Western blot for phospho-EGFR"
        detection_method: "chemiluminescence"
        antibodies_used:
          - "anti-pEGFR Y1068 (Cell Signaling #3777)"
          - "anti-total EGFR (Cell Signaling #4267)"
        normalization_method: "total protein normalization"