Journal: Scientific Reports

Article Title: LIM Protein Ajuba associates with the RPA complex through direct cell cycle-dependent interaction with the RPA70 subunit

doi: 10.1038/s41598-018-27919-8

Figure Lengend Snippet: Ajuba is in association with the RPA complex, and dissociates upon hydroxyurea treatment. ( A ) IP-Western on immunoprecipitations with Ajuba serum from HTC75 total extracts. The blots were probed with the indicated RPA antibodies. ( B ) IP-Western using the indicated RPA antibodies for immunoprecitations, and blotted for Ajuba. ( C ) IP-Western using anti-Ajuba antibodies for immunoprecipitations, and blotted for RPA32. Total extracts shown on the left panel. Cells were untreated (control), treated with 2 mM hydroxyurea (HU) for 24 hr, or synchronized by double Thymidine block and released for 2 hr. See also Figures , and for data with IMR90 cells. Panels shown are cropped from full-length blots shown in Supplemental Figure .

Article Snippet: HTC75 cells were cultured in DMEM (Cellgro) with 10% BCS (HyClone), 1% penicillin and streptomycin (Cellgro) and 1% L-glutamine (Gibco).

Techniques: Western Blot, Control, Blocking Assay

Journal: Scientific Reports

Article Title: LIM Protein Ajuba associates with the RPA complex through direct cell cycle-dependent interaction with the RPA70 subunit

doi: 10.1038/s41598-018-27919-8

Figure Lengend Snippet: Ajuba nuclear localization is increased during S phase. HTC75 cells were synchronized to G1/S border with double thymidine block and released into S phase, with cells processed for IF at the indicated time points. (Top) Immunofluorescence of Ajuba in unsynchronized and synchronized cells. (Bottom) Quantitation of cells positive for nuclear Ajuba at each time point (n = 100).

Article Snippet: HTC75 cells were cultured in DMEM (Cellgro) with 10% BCS (HyClone), 1% penicillin and streptomycin (Cellgro) and 1% L-glutamine (Gibco).

Techniques: Blocking Assay, Immunofluorescence, Quantitation Assay

Journal: Scientific Reports

Article Title: LIM Protein Ajuba associates with the RPA complex through direct cell cycle-dependent interaction with the RPA70 subunit

doi: 10.1038/s41598-018-27919-8

Figure Lengend Snippet: Nuclear Ajuba is found primarily in BrdU-positive cells. HTC75 cells were synchronized, then released in BrdU-containing medium. The BrdU was washed off after 1 hour. Cells were co-stained for Ajuba and BrdU at the indicated time points. The percent of nuclei with >3 foci of co-localization is shown (bottom right).

Article Snippet: HTC75 cells were cultured in DMEM (Cellgro) with 10% BCS (HyClone), 1% penicillin and streptomycin (Cellgro) and 1% L-glutamine (Gibco).

Techniques: Staining

Journal: Scientific Reports

Article Title: LIM Protein Ajuba associates with the RPA complex through direct cell cycle-dependent interaction with the RPA70 subunit

doi: 10.1038/s41598-018-27919-8

Figure Lengend Snippet: Increase of Ajuba-RPA70 co-localization in the nucleus during S phase. HTC75 cells were synchronized to G1/S border with double thymidine block and released into S phase with cells processed for IF at the indicated time points. (Top) Co-immunofluorescence of Ajuba and RPA70 in unsynchronized and synchronized cells. Arrowheads point to sites of co-localization (Bottom) Quantitation of cells exhibiting >3 foci of Ajuba-RPA70 co-localization in the nucleus at each time point (n = 100).

Article Snippet: HTC75 cells were cultured in DMEM (Cellgro) with 10% BCS (HyClone), 1% penicillin and streptomycin (Cellgro) and 1% L-glutamine (Gibco).

Techniques: Blocking Assay, Immunofluorescence, Quantitation Assay

Journal: Scientific Reports

Article Title: LIM Protein Ajuba associates with the RPA complex through direct cell cycle-dependent interaction with the RPA70 subunit

doi: 10.1038/s41598-018-27919-8

Figure Lengend Snippet: Ajuba nuclear localization is reduced upon HU treatment. (Left) Co-immunofluorescence of Ajuba (FITC) and RPA70 (TRITC) in untreated and HU treated HTC75 cells. (Right) Quantification of cells exhibiting strong nuclear localization with and without HU treatment (n = 100, three independent experiments).

Article Snippet: HTC75 cells were cultured in DMEM (Cellgro) with 10% BCS (HyClone), 1% penicillin and streptomycin (Cellgro) and 1% L-glutamine (Gibco).

Techniques: Immunofluorescence

Journal: Cell reports

Article Title: Shwachman-Diamond Syndrome Protein SBDS Maintains Human Telomeres by Regulating Telomerase Recruitment

doi: 10.1016/j.celrep.2018.01.057

Figure Lengend Snippet: (A) The inducible SBDS-knockout HeLa cells were harvested 6 days after doxycycline treatment (1 μg/mL) for western blot analysis. Endogenous SBDS protein levels were quantified using the Odyssey Infrared Imaging System and normalized against the GAPDH loading control.

Article Snippet: HEK293T, HTC75, and HeLa cells were cultured in DMEM (Corning Life Sciences) supplemented with 10% fetal bovine serum (Gibco) and 100 U/mL streptomycin/penicillin (Gibco) at 37°C in a 5% CO 2 incubator.

Techniques: Knock-Out, Western Blot, Imaging, Control