A re-evaluation of LINE-1 ORF2 expression in LNCaP prostate cancer cells
Abstract Background We previously examined expression of Long Interspersed Element-1 (LINE-1) in a variety of prostate cancer cells including hormone-dependent LNCaP cells. These studies demonstrated expression and sub-cellular localization of LINE-1 proteins, ORF1p, with antibody 4H1, and ORF2p, wi...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-12-01
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| Series: | Mobile DNA |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13100-019-0196-x |
| _version_ | 1819057496189304832 |
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| author | Erica M. Briggs Corrado Spadafora Susan K. Logan |
| author_facet | Erica M. Briggs Corrado Spadafora Susan K. Logan |
| author_sort | Erica M. Briggs |
| collection | DOAJ |
| description | Abstract Background We previously examined expression of Long Interspersed Element-1 (LINE-1) in a variety of prostate cancer cells including hormone-dependent LNCaP cells. These studies demonstrated expression and sub-cellular localization of LINE-1 proteins, ORF1p, with antibody 4H1, and ORF2p, with antibody chA1-L1. Results Here we conduct immunoprecipitation/mass spectrometry analysis using chA1-L1 antibody against ORF2p in LNCaP cells. Our results indicate that antigens detected by the antibody include the transcriptional regulator BCLAF1. We show that chA1-L1 recognizes BCLAF1 using siRNA knockdown and overexpression of a tagged BCLAF1. We also show that chA1-L1 antibody recognizes ORF2p in HEK293 cells overexpressing LINE-1. Further, analysis of ORF2p (chA1-L1) and BCLAF1 foci using immunofluorescence in LNCaP cells showed significant colocalization. Conclusions Overall, our findings indicate that chA1-L1 antibody recognizes both BCLAF1 and ORF2p but the majority of antigen recognized in LNCaP cells is BCLAF1. |
| first_indexed | 2024-12-21T13:40:14Z |
| format | Article |
| id | doaj.art-5d8064dcde444dbd8f4d1b582f9119c2 |
| institution | Directory Open Access Journal |
| issn | 1759-8753 |
| language | English |
| last_indexed | 2024-12-21T13:40:14Z |
| publishDate | 2019-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Mobile DNA |
| spelling | doaj.art-5d8064dcde444dbd8f4d1b582f9119c22022-12-21T19:02:03ZengBMCMobile DNA1759-87532019-12-011011610.1186/s13100-019-0196-xA re-evaluation of LINE-1 ORF2 expression in LNCaP prostate cancer cellsErica M. Briggs0Corrado Spadafora1Susan K. Logan2Departments of Biochemistry and Molecular Pharmacology, New York University School of Medicine, Alexandria Center for Life SciencesInstitute of Translational Pharmacology, National Research CouncilDepartments of Biochemistry and Molecular Pharmacology, New York University School of Medicine, Alexandria Center for Life SciencesAbstract Background We previously examined expression of Long Interspersed Element-1 (LINE-1) in a variety of prostate cancer cells including hormone-dependent LNCaP cells. These studies demonstrated expression and sub-cellular localization of LINE-1 proteins, ORF1p, with antibody 4H1, and ORF2p, with antibody chA1-L1. Results Here we conduct immunoprecipitation/mass spectrometry analysis using chA1-L1 antibody against ORF2p in LNCaP cells. Our results indicate that antigens detected by the antibody include the transcriptional regulator BCLAF1. We show that chA1-L1 recognizes BCLAF1 using siRNA knockdown and overexpression of a tagged BCLAF1. We also show that chA1-L1 antibody recognizes ORF2p in HEK293 cells overexpressing LINE-1. Further, analysis of ORF2p (chA1-L1) and BCLAF1 foci using immunofluorescence in LNCaP cells showed significant colocalization. Conclusions Overall, our findings indicate that chA1-L1 antibody recognizes both BCLAF1 and ORF2p but the majority of antigen recognized in LNCaP cells is BCLAF1.https://doi.org/10.1186/s13100-019-0196-xBCLAF1ORF2ORF2 antibodyLINE-1Prostate cancer |
| spellingShingle | Erica M. Briggs Corrado Spadafora Susan K. Logan A re-evaluation of LINE-1 ORF2 expression in LNCaP prostate cancer cells Mobile DNA BCLAF1 ORF2 ORF2 antibody LINE-1 Prostate cancer |
| title | A re-evaluation of LINE-1 ORF2 expression in LNCaP prostate cancer cells |
| title_full | A re-evaluation of LINE-1 ORF2 expression in LNCaP prostate cancer cells |
| title_fullStr | A re-evaluation of LINE-1 ORF2 expression in LNCaP prostate cancer cells |
| title_full_unstemmed | A re-evaluation of LINE-1 ORF2 expression in LNCaP prostate cancer cells |
| title_short | A re-evaluation of LINE-1 ORF2 expression in LNCaP prostate cancer cells |
| title_sort | re evaluation of line 1 orf2 expression in lncap prostate cancer cells |
| topic | BCLAF1 ORF2 ORF2 antibody LINE-1 Prostate cancer |
| url | https://doi.org/10.1186/s13100-019-0196-x |
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