Protocol detail

Quantifying Gene Expression in Mouse Lung and Calu-3 Cells

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RNA was isolated from whole mouse lung tissue or Calu-3 cells using the RNeasy Spin column kit (Qiagen #75144) then reverse transcribed using the cDNA Reverse Transcription Kit (Applied Biosystems #4308228) as previously described^{46 (link)}. Gene expression was reported as fold change over WT-saline condition normalized to GAPDH in mouse lung tissues or OAZ1 for Calu-3 cells and calculated using the double delta CT (2∆∆CT) method as previously described^{7 (link),8 (link),46 (link)}.

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Sweeter J.M., Kudrna K., Hunt K., Thomes P., Dickey B.F., Brody S.L, & Dickinson J.D. (2021). Autophagy of mucin granules contributes to resolution of airway mucous metaplasia. Scientific Reports, 11, 13037.

Publication 2021

RNeasy Spin column kit cDNA Reverse Transcription Kit

Cdna Cells Gapdh Gene expression Lung Mouse Reverse transcription Saline Tissues

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Variable analysis

independent variables

Whole mouse lung tissue
Calu-3 cells

dependent variables

Gene expression reported as fold change over WT-saline condition normalized to GAPDH in mouse lung tissues
Gene expression reported as fold change over WT-saline condition normalized to OAZ1 for Calu-3 cells

control variables

WT-saline condition

controls

Positive control: Not specified
Negative control: WT-saline condition

Annotations

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