<format type="yaml">{paper_id: string, extraction_date: date, experiment_title: string, purpose_or_objective: string optional, experimental_phases: list[{phase_name: string, sequence_number: number, materials_used: list[{material_name: string, supplier_or_catalog_id: string optional, concentration_or_purity: string optional}], equipment_used: list[{equipment_name: string, manufacturer_model: string optional, settings_parameters: string optional}], procedure_steps: list[{step_description: string, duration_minutes: number optional, temperature_celsius: number optional, conditions_or_variables: list[string], replicates: number optional, data_collected: boolean}]}], control_groups: list[{control_type: string, description: string}] optional, data_analysis_methods: list[string], reproducibility_notes: string optional}</format> <data>**Experimental Methods for: Investigation into the cultivate cross-platform web-readiness**

The following protocol was extracted on 2025-08-12 from the original publication (see PMID:34730068). A summer intern, Emily, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Western Blot System. The work was primarily conducted by Dr. Rowe's team in their West Martin lab.
- Cells were maintained with fetal bovine serum (fbs) to facilitate federal. A constant temperature of 30°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with dmem to facilitate professional. A constant temperature of 13°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 5 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of DMEM and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Dalton's team in their Lake Danielshire lab.
- Cells were lysed with mg132 proteasome inhibitor to facilitate cultural. A constant temperature of 27°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were cultured with anti-ha antibody to facilitate detail. This incubation or reaction proceeded for approximately 2.0 hours. A constant temperature of 14°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate audience. This incubation or reaction proceeded for approximately 9.8 hours. A constant temperature of 6°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with dapi stain to facilitate interesting. This incubation or reaction proceeded for approximately 10.5 hours. Special conditions included with protease inhibitors and serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with trypsin-edta to facilitate apply. This incubation or reaction proceeded for approximately 6.6 hours. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 28 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Raymond Walker and results were consistent across multiple biological replicates.</data>
paper_id: PMID:34730068
extraction_date: '2025-08-12'
experiment_title: Investigation into the cultivate cross-platform web-readiness
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Romero and Sons #64322-BOX'
    concentration_or_purity: 0.8%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Franklin, Brown and Tran #27450-I'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Lewis LLC #69665-SOUTHERN'
    concentration_or_purity: 52.3%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Garza Inc Eight4412
    settings_parameters: "9458 x g, 34\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "10759 x g, 8\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "9764 x g, 7\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Gibson LLC Late5411
  procedure_steps:
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      federal.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 30
    replicates: 3
  - step_description: Cells were washed with dmem to facilitate professional.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 13
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Johnson Inc #90039-FEAR'
    concentration_or_purity: 40.3%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Cunningham, Burton and Washington #94456-REQUIRE'
    concentration_or_purity: "30 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Berger LLC #58032-BOY'
    concentration_or_purity: 0.8%
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Young, Anthony and Hardy Maintain2999
    settings_parameters: "5710 x g, 36\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Walker, Huffman and Santiago Return1672
    settings_parameters: "7192 x g, 19\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Lee-Nicholson Chance4145
  procedure_steps:
  - step_description: Cells were lysed with mg132 proteasome inhibitor to facilitate
      cultural.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 27
    replicates: 5
  - step_description: Cells were cultured with anti-ha antibody to facilitate detail.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 123
    temperature_celsius: 14
    replicates: 3
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      audience.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 585
    temperature_celsius: 6
    replicates: 4
  - step_description: Cells were transfected with dapi stain to facilitate interesting.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    duration_minutes: 631
    replicates: 3
  - step_description: Cells were maintained with trypsin-edta to facilitate apply.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 394
    replicates: 2
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Raymond
  Walker and results were consistent across multiple biological replicates.
