csii pref1a mcherry3×nls  (Addgene inc)

Journal: bioRxiv

Article Title: Live dynamics of induced cell-cell fusion between mitotic and interphasic cells

doi: 10.64898/2026.01.27.700572

Figure Lengend Snippet: (A and B) Analysis of nuclear pore formation in control telophase cells and cells fixed at different time points after cell-cell fusion and respective fluorescence intensity quantification. Magenta square shows nuclear envelope membranes on condensed chromosomes but no nuclear pore protein accumulation. Control, n=19 cells; 20 min, n=20 cells; 30 min, n=22 cells; 40 min, n=27 cells; 50 min, n=32 cells and 60 min, n=26 cells. Statistics, non-parametric Mann-Whitney test. (C) Time-lapse of mCherry-NLS in control and mitotic fused cells shows efficient import of the NLS signal in control and fused interphase nucleus (cyan arrow), but not in mitotic fused nucleus (magenta arrows). Time is in h:min. Scale bar is 10μm. (D) Ratio of nuclear and cytoplasmic NLS signal of the examples shown in (C).

Article Snippet: The VSV-G (#12259), BFP and mCherry-NLS plasmids were obtained from Addgene.

Techniques: Control, Fluorescence, MANN-WHITNEY

Journal: Nucleic Acids Research

Article Title: PARP1 and PARP2 are dispensable for DNA repair by microhomology-mediated end-joining at double-ended DSBs

doi: 10.1093/nar/gkaf1437

Figure Lengend Snippet: PARP inhibitors increase MMEJ at ISceI-mediated deDSBs. ( A, B ) Schematic ( A ) and experimental timeline ( B ) of MMEJ reporter used in panels (C)–(F), Figs and , , , and E, and , and and . Cutting the reporter with ISceI and repair by microhomology annealing to an in-frame, functional mCherry gene. Flow cytometry was used to identify the mCherry + cells (MMEJ+) within the BFP+ (ISceI+) population. ( C–E ) MMEJ quantification using reporter and timeline from panel (A) and (B) in HT1080 cells. Values are normalized to DMSO. Drugs used were olaparib (5 µM or as indicated), DNA-PKcsi (NU7441, 1 µM), Polθi (ART558 10 µM), niraparib (2.5 µM), rucaparib (2.5 µM), and talazoparib (2.5 µM). ( F ) MMEJ quantification in HT1080 parental cells compared to isogenic POLQ -KO cells following olaparib treatment. Values are normalized to wild-type DMSO. Statistical analyses for panels (C)–(F): Data represent three independent experiments, each the average of three technical replicates. Data are mean ± SEM. Statistical test, one way ANOVA with multiple comparison correction. ns: nonsignificant, * P <.05, ** P <.01, *** P <.001, **** P <.0001.

Article Snippet: The PARP2 plasmid used to complement PARP2 -KO clones was a generous gift from the Karolin Luger lab and was derived by cloning the human PARP2 coding region into plasmid mCherry C1 (obtained from Dyche Mullins [ ], Addgene #58476).

Techniques: Functional Assay, Flow Cytometry, Comparison

Journal: Nucleic Acids Research

Article Title: PARP1 and PARP2 are dispensable for DNA repair by microhomology-mediated end-joining at double-ended DSBs

doi: 10.1093/nar/gkaf1437

Figure Lengend Snippet: PARPi-dependent MMEJ increase does not occur at Cas9-induced DSBs. ( A, B ) Schematic ( A ) and experimental timeline ( B ) of MMEJ reporter used in panels (C)–(E), Figs and . Repair of the Cas9-induced cut with microhomology annealing leads to an in-frame, functional mCherry gene. Flow cytometry was used to identify the mCherry + cells (MMEJ+) within the GFP+ (Cas9+) population. ( C–E ) MMEJ quantification using the reporter and timeline from panels (A) and (B) in HT1080 cells. Values are normalized to DMSO. Drugs used were olaparib (5 µM or indicated dose), DNA-PKcsi (NU7441, 1 µM), Polθi (ART558, 10 µM). ( F ) HR quantification using the DR-GFP reporter in HT1080 cells after an ISceI- or Cas9-induced DSB, ± olaparib treatment (5 µM). Statistical analyses ( C–F ): Data represent three (D–F) or four ( C ) independent experiments, each the average of three technical replicates. Data are mean ± SEM. Statistical test, one way ANOVA with multiple comparison correction. ns: nonsignificant, * P <.05, ** P <.01, *** P <.001, **** P <.0001.

Article Snippet: The PARP2 plasmid used to complement PARP2 -KO clones was a generous gift from the Karolin Luger lab and was derived by cloning the human PARP2 coding region into plasmid mCherry C1 (obtained from Dyche Mullins [ ], Addgene #58476).

Techniques: Functional Assay, Flow Cytometry, Comparison

Journal: bioRxiv

Article Title: Dual Regulatory Role of Nuclear FNBP4 in Actin Binding and Formin FMN1 Inhibition

doi: 10.64898/2026.01.05.697755

Figure Lengend Snippet: (A) Cells were transfected with the pmCherry-C1 actin-3×NLS P2A mCherry construct. After transfection, the cells were fixed with 3.7% paraformaldehyde and subjected to immunofluorescence staining for FNBP4. FNBP4 was detected using an anti-FNBP4 antibody, and nuclei were counterstained with DAPI. The representative image shows a single optical section from the middle of the Z-stack. Scale bar: 20 µm. (B) Quantification of FNBP4 and actin colocalization using Manders’ Colocalization Coefficient. The scatter dot plot shows the distribution of data points along with standard deviations.

Article Snippet: For transfection studies, the pmCherry-C1 actin-3×NLS P2A mCherry plasmid was obtained from Addgene (RRID:Addgene_58475; deposited by Dyche Mullins).

Techniques: Transfection, Construct, Immunofluorescence, Staining