The smoke from 15 lit cigarettes (Sichuan China Tobacco Industry Co., Ltd.) was slowly inhaled into a 50 ml syringe, and then injected it into DMEM pre-heated in a 37˚C water bath (38 (link)). The pH of DMEM was adjusted to 7.4 and sterilized using a 0.22-µm filter (EMD Millipore) and stored at -80˚C. Serum-free DMEM was used to dilute 100% CSE to the required CSE concentrations (5, 10 and 20%). Cells were treated with vitamin D3 (250, 500 or 1,000 nM; Sigma-Aldrich; Merck KGaA) or vehicle (0.1% ethanol) for 30 min prior to 24 h treatment at 37˚C with CSE and vitamin D3 or vehicle, after shRNA transfection (when it was required) (26 (link)).
Protocol detail
Cigarette Smoke Extract and Vitamin D in Cell Culture
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Zhejiang Chinese Medical University, Fourth Hospital of Inner Mongolia
Top 5 similar protocols
Variable analysis
independent variables
- Cigarette smoke exposure (CSE) concentration (5%, 10%, and 20%)
- Vitamin D3 concentration (250 nM, 500 nM, and 1,000 nM)
- Cellular response to CSE and vitamin D3 treatment
- PH of DMEM adjusted to 7.4
- DMEM pre-heated in a 37°C water bath
- DMEM sterilized using a 0.22-µm filter
- DMEM stored at -80°C
- Serum-free DMEM used to dilute 100% CSE
- Cells treated with vehicle (0.1% ethanol) for 30 min prior to 24 h treatment
- Not explicitly mentioned
- Cells treated with vehicle (0.1% ethanol) for 30 min prior to 24 h treatment
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!