Stable Luciferase-Expressing KRAS Mutant Cell Lines

CMT167 cells were stably transfected with firefly luciferase, as previously described (24 (link)). Luciferase-expressing LLC cells were purchased from Caliper Life Sciences (LL/2-luc-M38). The CMT167 cells harbor a Kras^G12V mutation, and the LLC cells harbor a Kras^G12C mutation (24 (link)). Both cell lines were maintained in DMEM with 4.5 g/l glucose (no. 10–017-CV; Corning) containing 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 500 μg/ml G418 at 37°C in a humidified 5% CO₂ atmosphere. Cells were periodically tested for mycoplasma infection, maintained as frozen stocks, and cultured for only 2–4 wk before use in experiments. Authentication of cell lines based on morphology, growth curve analysis, and metastatic phenotype was performed regularly. Human NSCLC cell lines were maintained in RPMI 1640 (no. 10–040-CV; Corning) containing 10% FBS at 37°C in a humidified 5% CO₂ atmosphere. All were KRAS mutant.

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.

Johnson A.M., Bullock B.L., Neuwelt A.J., Poczobutt J.M., Kaspar R.E., Li H.Y., Kwak J.W., Hopp K., Weiser-Evans M.C., Heasley L.E., Schenk E.L., Clambey E.T, & Nemenoff R.A. (2020). Cancer Cell–Intrinsic Expression of MHC Class II Regulates the Immune Microenvironment and Response to Anti–PD-1 Therapy in Lung Adenocarcinoma. Journal of immunology (Baltimore, Md. : 1950), 204(8), 2295-2307.

Publication 2020

LL/2-luc-M38 DMEM with 4.5 g/l glucose RPMI 1640

Atmosphere Cell lines Cell lines authentication Cells Firefly luciferase Frozen G418 Glucose Human Kras Luciferase Mutation Mycoplasma infection Nsclc Penicillin Phenotype Streptomycin

Corresponding Organization : University of Colorado Anschutz Medical Campus

Top 5 similar protocols

Variable analysis

independent variables

Stable transfection of CMT167 cells with firefly luciferase
Kras mutation status in cell lines (CMT167 cells harbor Kras^G12V, LLC cells harbor Kras^G12C)

dependent variables

Luciferase expression in CMT167 and LLC cells
Cell morphology, growth curve, and metastatic phenotype

control variables

Cell culture conditions (DMEM with 4.5 g/l glucose, 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, 500 μg/ml G418, 37°C, 5% CO2 atmosphere)
Mycoplasma infection testing
Cell line authentication based on morphology, growth curve analysis, and metastatic phenotype

positive controls

Luciferase-expressing LLC cells purchased from Caliper Life Sciences (LL/2-luc-M38)

negative controls

None specified

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.

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?

Revolutionizing how scientists
search and build protocols!