3xflag  (Addgene inc)

Journal: NAR Cancer

Article Title: Effects of hydrazone-based G-quadruplex ligands on FANCJ/BRIP1 -depleted cancer cells and a Caenorhabditis elegans dog-1 −/− strain

doi: 10.1093/narcan/zcaf004

Figure Lengend Snippet: Analysis of G4 focus formation in FANCJ -KO HeLa cells. ( A ) Immunofluorescence showing BG4 foci in the indicated cell lines. HeLa cells were treated for 24 h with PDS (10 μM). Scale bar, 5 μm. ( B ) Fold increase of the BG4 CTF, detected per nucleus in the indicated cell lines and conditions. The chart reports medians (lines), mean (cross), 50% (box), and 90% (whiskers) of the datasets and outliers (circles). A total of 100 cells were analysed per condition in three biological independent experiments. Statistical analyses were performed using a non-parametrical Mann–Whitney U -test. Two-tailed P -values were calculated (**** P < .0001).

Article Snippet: Recombinant 6xHis/3xFLAG-tagged BG4 was produced in Escherichia coli using pSANG10-3F-BG4 (Addgene; #55756) as follows: BL21(DE3) competent cells were transformed with pSANG10-3F plasmid expressing BG4 scFv anti-G4 antibody.

Techniques: Immunofluorescence, MANN-WHITNEY, Two Tailed Test

Journal: NAR Cancer

Article Title: Effects of hydrazone-based G-quadruplex ligands on FANCJ/BRIP1 -depleted cancer cells and a Caenorhabditis elegans dog-1 −/− strain

doi: 10.1093/narcan/zcaf004

Figure Lengend Snippet: Quantitative analysis of G4 stabilization in FANCJ -KO cells by different G4 binders. ( A ) Immunofluorescence representative images showing BG4 foci in the indicated cell lines after treatment with PDS (500 nM), FIM (500 nM), and FIM-15 (250 nM) for 24 h. Scale bar, 5 μm. ( B ) Fold increase of the BG4 CTF, detected per nucleus in the indicated cell lines and conditions. The chart reports medians (lines), mean (cross), 50% (box), and 90% (whiskers) of the datasets and outliers (circles). A total of 100 cells were analysed per condition in three biological independent experiments. Statistical analyses were performed using a non-parametrical Mann–Whitney U -test. Calculated P -values were indicated as * P < .05; **** P < .0001.

Article Snippet: Recombinant 6xHis/3xFLAG-tagged BG4 was produced in Escherichia coli using pSANG10-3F-BG4 (Addgene; #55756) as follows: BL21(DE3) competent cells were transformed with pSANG10-3F plasmid expressing BG4 scFv anti-G4 antibody.

Techniques: Immunofluorescence, MANN-WHITNEY

Journal: NAR Cancer

Article Title: Effects of hydrazone-based G-quadruplex ligands on FANCJ/BRIP1 -depleted cancer cells and a Caenorhabditis elegans dog-1 −/− strain

doi: 10.1093/narcan/zcaf004

Figure Lengend Snippet: Detection of G4 foci in the gonadal nuclei of C. elegans dog-1 − / − . The indicated worm strains were subjected to a 24-h treatment with FIM-15 (20 μM). Nuclei were co-stained with the BG4 antibody and DAPI. Scale bar, 20 μm. Insets, indicated by boxes, show sections of the main images corresponding to the mitotic zone and pachytene stage nuclei. Scale bar, 5 μm.

Article Snippet: Recombinant 6xHis/3xFLAG-tagged BG4 was produced in Escherichia coli using pSANG10-3F-BG4 (Addgene; #55756) as follows: BL21(DE3) competent cells were transformed with pSANG10-3F plasmid expressing BG4 scFv anti-G4 antibody.

Techniques: Staining