Western Blot Analysis of Cellular Proteins

As previously described (24 (link), 25 (link)), proteins were extracted and normalized to at least 1500μg/ml. Proteins were subsequently analyzed on SDS-PAGE gels (8% or 10%, as appropriate) and transferred to polyvinylidene difluoride (PVDF) membranes for western blotting (24 (link), 25 (link)).
Primary antibodies that were utilized in this study are: SV40 LTAg mouse monoclonal (#554149, 1:1000, BD Biosciences), DNMT1 rabbit polyclonal (#5032, 1:1000, Cell Signaling Technology), HA-11 mouse monoclonal (#14921901, 1:500, Covance/Lab Corporation), CD71 mouse monoclonal (#sc-51829, 1:250, Santa Cruz Biotechnology), FTH1 rabbit polyclonal (#3998, 1:500, Cell Signaling Technology), pan-Actin rabbit polyclonal (#4968, 1:1000, Cell Signaling Technology), PARP rabbit polyclonal (#9542, 1:1000, Cell Signaling Technology), PAX-8 rabbit polyclonal (#10336-1-AP, 1:1000, Proteintech), Claudin-1 rabbit polyclonal (#187362, 1:10,000, Invitrogen), BAK rabbit monoclonal (#12105, 1: 1000, Cell Signaling Technology), Bcl-2 rabbit monoclonal (#2570, 1:1000, Cell Signaling Technology), Occludin-1 mouse monoclonal (#611090, 1:250, BD Biosciences), Cyclin D1 rabbit polyclonal (#sc-718, 1:1000, Santa Cruz Biotechnology), and Cyclin E mouse monoclonal (#sc-247, 1:500, Santa Cruz Biotechnology).

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Chhabra R., Guergues J., Wohlfahrt J., Rockfield S., Espinoza Gonzalez P., Rego S., Park M.A., Berglund A.E., Stevens SM J.r, & Nanjundan M. (2023). Deregulated expression of the 14q32 miRNA cluster in clear cell renal cancer cells. Frontiers in Oncology, 13, 1048419.

Publication 2023

SV40 LTAg mouse monoclonal FTH1 rabbit polyclonal pan-Actin rabbit polyclonal PARP rabbit polyclonal Cyclin D1 rabbit polyclonal

Actin Antibodies Bcl 2 Cd71 Claudin 1 Cyclin d1 Cyclin e Dnmt1 Gels Membranes Mouse Occludin Polyvinylidene difluoride Proteins Rabbit Sds page Sv40

Corresponding Organization : University of South Florida

Other organizations : St. Jude Children's Research Hospital, Moffitt Cancer Center

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of SV40 LTAg, DNMT1, HA-11, CD71, FTH1, pan-Actin, PARP, PAX-8, Claudin-1, BAK, Bcl-2, Occludin-1, Cyclin D1, Cyclin E

control variables

None explicitly mentioned

controls

Positive controls: none mentioned
Negative controls: none mentioned

Annotations

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