Journal: Nature Communications

Article Title: Structural mechanisms for centrosomal recruitment and organization of the microtubule nucleator γ-TuRC

doi: 10.1038/s41467-025-57729-2

Figure Lengend Snippet: a Representative immunofluorescence images of RPE1 cells in different cell cycle stages stained against γ-tubulin (yellow) and HAUS1-SNAP-S with anti-Strep antibodies (HAUS1-Strep, magenta). DNA was visualized with DAPI (blue). Green arrows indicate the centrioles shown in the close-up view. b , c Quantification of fluorescence intensity at centrosomes for the indicated proteins shown in a) and Supplementary Fig. . n = 125 (interphase), 132 (prophase), 121 (prometaphase), 125 (metaphase), 162 (anaphase), 128 (telophase), 126 (+ STLC), and 117 (+ STLC&BI2536) centrioles were analyzed for HAUS4 intensity. n = 108 (interphase), 127 (prophase), 121 (prometaphase), 126 (metaphase), 121 (anaphase), 122 (telophase), 120 (+ STLC), and 120 (+ STLC&BI2536) centrioles were analyzed for γ-tubulin intensity. n = 127 (interphase), 118 (prophase), 130 (prometaphase), 123 (metaphase), 121 (anaphase), 127 (telophase), 131 (+ STLC), and 119 (+ STLC&BI2536) centrioles were analyzed for POC5 intensity. d Immunoblot showing protein levels of HAUS4 and γ-tubulin in wild-type control, POC5 -/- , POC5 and POC5 mut cells. e Quantification of relative HAUS4 and γ-tubulin levels in ( d ) (normalized to actin). f Representative immunoblots of POC5 -/- cells expressing POC5 or POC5 mut , treated with 50 µg/ml CHX for the indicated durations, using anti-HAUS4, anti-γ-tubulin and anti-GAPDH (loading control) antibodies. g ) Quantification of panel f ). Relative protein levels were normalized to protein levels at time 0. Linear regression was performed; R 2 values are provided in the Source Data. h Representative images of wild-type control, POC5 -/- , POC5, and POC5 mut cells in prophase and metaphase stained against the centrosomal marker γ-tubulin (green). DNA was visualized with DAPI. Green arrows indicate misaligned chromosomes. i Quantification of cells with amplified γ-tubulin foci and misaligned chromosomes for h). N = 139 (control), 136 ( POC5 -/- ), 129 ( POC5 ), and 137 ( POC5 mut ) cells were analyzed for amplified γ-tubulin foci. n = 135 (control), 133 ( POC5 -/- ), 125 ( POC5 ) and 131 ( POC5 mut ) cells were analyzed for misaligned chromosomes. ( a ), ( h ), scale bars: 10 µm, magnification scale bars: 1 µm; in ( b , c , e , i ), data are presented as mean ± SD, and all statistics were derived from ordinary one-way ANOVA analysis of N = 3 biologically independent experiments. Source data are provided as a Source Data file.

Article Snippet: To test the function of PLK1 in mitosis, cells were treated with 5 μM of STLC 4 hr, followed by 2 h of 50 nM of BI2536 (Cell signaling) treatment.

Techniques: Immunofluorescence, Staining, Fluorescence, Western Blot, Control, Expressing, Marker, Amplification, Derivative Assay

Journal: Viruses

Article Title: Development of Oncolytic Vectors Based on Human Adenovirus Type 6 for Cancer Treatment.

doi: 10.3390/v15010182

Figure Lengend Snippet: Figure 1. Construction of Ad6-hT-GM. pShuttle is based on pBR322 and contains the left part of the Ad6 genome from LITR to 1773 bp (shown in yellow) with hTERT promoter (shown in purple) replacing the native E1A promoter and the right part, from 27,576 bp to RITR (shown in green) with the GM-CSF transgene inserted instead of E3-6.7K and E3-gp19K genes (shown in cyan). pShuttle was linearized by amplification with primers containing 15 nucleotide homology arms (shown in orange). The Ad6 genome was digested with ClaI and PacI, and recombination between truncated genome and linearized pShuttle was performed by the In-Fusion kit. Ad6-hT-GM recombinant virus genome was then retrieved from the plasmid with the AsiSI. Expanded regions indicate modified Ad6-hT-GM genome regions.

Article Snippet: After that, the native E1A promoter (360–559 bp) was substituted by hTERT promoter (from pAdVTERT plasmid #26744, Addgene) using primers TERT-pShuttle-for and TERT-pShuttle-rev (Table S1) for the amplification of hTERT and pShuttle-TERT-for and pShuttle-TERT-rev (Table S1) for the amplification of pShuttle resulting in pShuttle-hTE1A-AdITRs.

Techniques: Recombinant, Virus, Plasmid Preparation