PC-3, 22RV1, and DU145 human prostate-cancer cell lines were purchased from ATCC. We have previously verified that human metastatic prostate-cancer cells lack OLFM4 expression12 (link). The construction of vector-V5 tag, OLFM4-V5 tag, and truncated mutant OLFM4-Flag tag plasmids and their stable expression in plasmid cell clones have been previously described12 (link)16 (link). PC-3V (vector-transfected control) clone, PC-3W (full-length OLFM4-V5 tag-expressing) clone, and PC-3N (expressing a truncated mutant of OLFM4-Flag tag in which the olfactomedin domain is missing) clone cells were cultured in RPMI 1640 media containing 10% fetal bovine serum (FBS) until 90% confluent16 (link)28 (link). The pCMV-6-AC-GFP tag-vector and pCMV-6-OLFM4-GFP tag plasmids were purchased from Origene. Stably expressing OLFM4-green fluorescent protein (GFP) tag-expressing prostate-cancer cell clones (O) or vector-GFP tag-transfected control prostate-cancer cell clones (V) were established following previously described protocols12 (link)16 (link).
Pcmv 6 ac gfp tag vector
Manufactured by OriGene
The PCMV-6-AC-GFP tag-vector is a plasmid that contains the green fluorescent protein (GFP) gene under the control of the cytomegalovirus (CMV) promoter. This vector can be used for the expression and detection of proteins of interest in mammalian cells.
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2 protocols using pcmv 6 ac gfp tag vector
1
Prostate Cancer Cell Lines OLFM4 Expression
2
Generating OLFM4-expressing Prostate Cancer Cells
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We have previously verified that human prostate cancer cells lack OLFM4 expression.15 The pCMV‐6‐AC‐GFP tag‐vector and pCMV‐6‐OLFM4‐GFP tag plasmids were purchased from Origene (Rockville, MD). Stably expressing OLFM4‐GFP tag‐expressing prostate cancer cell clones (O) or vector‐GFP tag‐transfected control prostate cancer cell clones (V) were established following previously described protocols.15 , 33
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