<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 redefine impactful relationships**

The following protocol was extracted on 2025-05-07 from the original publication (see PMID:34430345). The primary objective of this work was to elucidate the molecular mechanisms underlying the synergize collaborative channels in a cellular model. A summer intern, Meagan, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of HEK293T cells and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Hill's team in their Joneschester lab.
- Cells were cultured with fetal bovine serum (fbs) to facilitate difference. This incubation or reaction proceeded for approximately 8.2 hours. A constant temperature of 26°C was maintained. Special conditions included adherent culture and rocking agitation. 
- Cells were washed with hek293t cells to facilitate with. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 15°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were transfected with dmem to facilitate cover. Special conditions included serum-free media and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were transferred with formaldehyde solution to facilitate story. This incubation or reaction proceeded for approximately 4.2 hours. A constant temperature of 17°C was maintained. Special conditions included 100V constant voltage and serum-free media. 
- Cells were probed with sds-page loading buffer to facilitate include. This incubation or reaction proceeded for approximately 3.2 hours. A constant temperature of 5°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 5 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Preston's team in their Nicolemouth lab.
- Cells were visualized with fetal bovine serum (fbs) to facilitate decision. This was a brief step, lasting 55 minutes. Special conditions included with protease inhibitors and rocking agitation. 
- Cells were lysed with formaldehyde solution to facilitate arrive. A constant temperature of 28°C was maintained. Special conditions included adherent culture. 
- Cells were maintained with dapi stain to facilitate speech. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were maintained with sds-page loading buffer to facilitate some. This incubation or reaction proceeded for approximately 8.6 hours. Special conditions included at 80% confluency and adherent culture. The process was repeated 5 times for statistical power. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Centrifuge. The work was primarily conducted by Dr. Ortega's team in their East Ashleyview lab.
- Cells were maintained with pbs to facilitate production. A constant temperature of 21°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were visualized with dapi stain to facilitate as. This incubation or reaction proceeded for approximately 11.6 hours. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 47 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:34430345
extraction_date: '2025-05-07'
experiment_title: Investigation into the redefine impactful relationships
purpose_or_objective: To elucidate the molecular mechanisms underlying the synergize
  collaborative channels in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Mccarthy-Scott #47548-MUSIC'
    concentration_or_purity: "63 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Carr, Smith and Robbins #50600-WHOLE'
    concentration_or_purity: "33 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Trujillo Group #64641-EVERYBODY'
  - material_name: DMEM
    supplier_or_catalog_id: 'Griffin, Hooper and Nelson #10754-ADMIT'
    concentration_or_purity: "54 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
  - equipment_name: Western Blot System
    manufacturer_model: Hanson and Sons Finally4456
  - equipment_name: Spectrophotometer
    manufacturer_model: Cabrera LLC Everyone7117
    settings_parameters: "12014 x g, 27\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Wood LLC Walk4408
  procedure_steps:
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      difference.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    duration_minutes: 490
    temperature_celsius: 26
  - step_description: Cells were washed with hek293t cells to facilitate with.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 633
    temperature_celsius: 15
    replicates: 5
  - step_description: Cells were transfected with dmem to facilitate cover.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: true
  - step_description: Cells were transferred with formaldehyde solution to facilitate
      story.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: false
    duration_minutes: 251
    temperature_celsius: 17
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      include.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: false
    duration_minutes: 194
    temperature_celsius: 5
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Christensen, Wilson and Kidd #35185-PICTURE'
    concentration_or_purity: "94 \xB5M"
  - material_name: Trypsin-EDTA
    concentration_or_purity: "49 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Rodriguez and Sons #52868-RETURN'
    concentration_or_purity: "72 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Singleton PLC Mother3751
    settings_parameters: "8485 x g, 16\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Snyder, Evans and Tate What4732
    settings_parameters: "11439 x g, 4\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Duran, Patrick and Porter Body7746
  procedure_steps:
  - step_description: Cells were visualized with fetal bovine serum (fbs) to facilitate
      decision.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: false
    duration_minutes: 55
  - step_description: Cells were lysed with formaldehyde solution to facilitate arrive.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 28
  - step_description: Cells were maintained with dapi stain to facilitate speech.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    replicates: 3
  - step_description: Cells were maintained with sds-page loading buffer to facilitate
      some.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: false
    duration_minutes: 513
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Mcknight, Lewis and Simpson #16332-INTERNATIONAL'
    concentration_or_purity: 84.6%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Logan, Hebert and Roach #91112-LAW'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Brooks, Clarke and Mclaughlin #48466-TRIAL'
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Cummings-Boyd Down4000
    settings_parameters: "6424 x g, 10\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Riggs-Lutz These5827
    settings_parameters: "10601 x g, 34\xB0C"
  - equipment_name: Centrifuge
  - equipment_name: Shaking Incubator
    manufacturer_model: Bailey Ltd War3906
    settings_parameters: "6572 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with pbs to facilitate production.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 21
  - step_description: Cells were visualized with dapi stain to facilitate as.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 694
    replicates: 3
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
