Journal: BMC research notes

Article Title: Centriolin interacts with HectD1 in a cell cycle dependent manner.

doi: 10.1186/s13104-023-06670-y

Figure Lengend Snippet: Fig. 1 Yeast two hybrid and immunoprecipitations reveal a direct interaction between centriolin and HectD1. A Schematic shows the N-terminal region (amino acids 1–250) of the centriolin protein that was fused to the DNA binding domain (DBD) of the pGBKT7 vector. This construct was used in a yeast two hybrid screen that revealed an interaction between centriolin and HectD1. B Hela cells were arrested in the indicated cell cycle stages and cell lysates were subjected to immunoprecipitation using antibodies for centriolin. Western blot was then performed on immunoprecipitates using either HectD1 antibodies or centriolin antibodies

Article Snippet: The following primary antibodies were used for immunofluorescence staining: centriolin (Thermo Fisher, cat # PA5-54219), α-tubulin (Invitrogen, cat. # 62204), and HectD1 (cat # sc-517169, Santa Cruz Biotechnology).

Techniques: Binding Assay, Plasmid Preparation, Construct, Two Hybrid Screening, Immunoprecipitation, Western Blot

Journal: BMC research notes

Article Title: Centriolin interacts with HectD1 in a cell cycle dependent manner.

doi: 10.1186/s13104-023-06670-y

Figure Lengend Snippet: Fig. 3 The expression of centriolin and HectD1 fluctuate throughout the cell cycle. Hela cells were arrested in the indicated cell cycle stages and western blot was performed on the cell lysates. HectD1 levels remain relatively constant in G0/G1, but fall during S phase, followed by a dramatic increase in mitosis. In contrast, centriolin displays a steady increase in expression from G0/G1 to S phase but drops in mitosis. Alpha tubulin was used as a loading control

Article Snippet: The following primary antibodies were used for immunofluorescence staining: centriolin (Thermo Fisher, cat # PA5-54219), α-tubulin (Invitrogen, cat. # 62204), and HectD1 (cat # sc-517169, Santa Cruz Biotechnology).

Techniques: Expressing, Western Blot, Control

Journal: BMC research notes

Article Title: Centriolin interacts with HectD1 in a cell cycle dependent manner.

doi: 10.1186/s13104-023-06670-y

Figure Lengend Snippet: Fig. 2 Centriolin and HectD1 co-localize at the spindle poles in mitosis. Immunofluorescence staining was performed on an asynchronous population of Hela cells. DAPI stain (blue, A) indicates the nuclei of individual cells and also reveals mitotic events. Antibodies against human HectD1 protein (green, B) and human centriolin protein (red, C) were used. Note that colocalization is present only in the cell undergoing mitosis (arrows), whereas no colocalization can be seen in interphase cells (asterisks). Colocalization is also shown in the Merge image (panel D), with quantitation in panel E (n = 3)

Article Snippet: The following primary antibodies were used for immunofluorescence staining: centriolin (Thermo Fisher, cat # PA5-54219), α-tubulin (Invitrogen, cat. # 62204), and HectD1 (cat # sc-517169, Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Staining, Quantitation Assay