CKAP4 Palmitoylation and Function in Lung Cancer

All antibodies, chemicals, and cell lines are shown in Tables S1-S3. A549, Calu-1, NCI-H292, and HCC4006 cells were authenticated in July 2022 using short tandem repeat analysis and mycoplasma testing (MycoStrip, InvivoGen). Target sequences for siRNA and shRNA assays are shown in Table S4. Primer sequences for quantitative polymerase chain reaction (PCR) are shown in Table S5. Anti-CKAP4 monoclonal antibodies were generated as previously described (19 (link)). Knockout (KO) cells were generated using the CRISPR/Cas9 system as previously described (26 (link)).
CKAP4^WT-HA and CKAP4^C100S-HA are wild-type (WT) CKAP4 and CKAP4 mutant in which cysteine at amino acid number 100 was changed to serine, respectively. CKAP4^C100S-HA is not palmitoylated. Both CKAP4s were tagged by HA at the C-terminal end.

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Nagoya A., Sada R., Kimura H., Yamamoto H., Morishita K., Miyoshi E., Morii E., Shintani Y, & Kikuchi A. (2023). CKAP4 is a potential exosomal biomarker and therapeutic target for lung cancer. Translational Lung Cancer Research, 12(3), 408-426.

Publication 2023

MycoStrip

Amino acid Anti antibodies Antibodies Assays Cell lines Cells Crispr Cysteine Mycoplasma Polymerase chain reaction Primer Serine Short tandem repeat Shrna Sirna

Corresponding Organization : Osaka University

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Variable analysis

independent variables

CKAP4^WT^-HA (wild-type CKAP4 tagged with HA)
CKAP4^C100S^-HA (CKAP4 mutant with cysteine 100 changed to serine, not palmitoylated, tagged with HA)

dependent variables

Not explicitly mentioned

control variables

Cell lines: A549, Calu-1, NCI-H292, and HCC4006
Cell line authentication and mycoplasma testing
SiRNA and shRNA target sequences
Primer sequences for quantitative PCR

controls

Positive control: Anti-CKAP4 monoclonal antibodies generated as previously described (19)
Negative control: Not explicitly mentioned

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