Journal: Scientific Reports

Article Title: Characterisation and use of a functional Gadd45g bacterial artificial chromosome

doi: 10.1038/s41598-018-35458-5

Figure Lengend Snippet: ( A ) Position of the mCherry reporter in the Gadd45g open reading frame of E15 BAC; ( B ) Detection of Cherry fluorescence in 11.5 dpc embryonic gonad. Green signal indicates location of PECAM. Blue is DAPI staining. ( C ) Cherry signal is detected in somatic cells of the gonad, which lack PECAM staining shown in ( D ). A few somatic cells (lacking PECAM) also lack Cherry signal (white arrows). ( E ) Lateral view of 10.5 dpc embryo showing Cherry fluorescence in developing neural tissue (forebrain (fb), midbrain (mb), hindbrain (hb)). ( F ) Dorsal view of same embryo reveals signal in neural tube (nt), trigeminal ganglion (tg) and facial ganglion (fg). ( G ) Section of embryo (in plane indicated by dotted line in ( E )) shows neural tube and dorsal root gangion (drg) fluorescence. Scale bar = 50 μm.

Article Snippet: A construct containing mCherry, nuclear localization signal and SV40pA terminator derived from Addgene plasmid GW1-Peredox-mCherry-NLS (#32381, a kind gift from Gary Yellen) was used to replace the rspL-neo cassette in a second round of transformation, restoring streptomycin resistance.

Techniques: Fluorescence, Staining

Journal: Free radical biology & medicine

Article Title: The cyclic nitroxide antioxidant 4-methoxy-TEMPO decreases mycobacterial burden in vivo through host and bacterial targets.

doi: 10.1016/j.freeradbiomed.2019.03.010

Figure Lengend Snippet: Fig. 5. Effects of MetT treatment on innate immune system. (A) qPCR quantification of proinflammatory gene expression in 5 dpi zebrafish embryos infected with M. marinum and treated with MetT. Genes are interleukin-1b (il1b), interleukin-8 (il8), matrix metalloprotease 9 (mmp9) and tumour necrosis factor a (tnfa). (B) Representative image of neutrophil recruitment to granulomas in 5 dpi durif/mpx homozygous mutants infected with M. marinum-tomato. White arrowheads indicate granulomas with mpx expressing neutrophil recruitment. Scale bar indicates 1 mm. (C) Representative images of 5 dpi zebrafish durif mutant embryos infected with M. marinum-tomato. (D) Quantification of bacterial burden by fluorescent pixel count in 5 dpi zebrafish durif mutant embryos. Bacterial fluorescence area data are expressed relative to the mean value for the heterozygous genotype group. (E) Quantification of bacterial burden by fluorescent pixel count in 5 dpi zebrafish embryos infected with ΔESX1 mutant M. marinum and treated with MetT as indicated. (F) Quantification of bacterial burden by fluorescent pixel count in 5 dpi zebrafish embryos treated MetT and dexamethasone as indicated. D-F each dot represents one embryo.

Article Snippet: To visualise disruption of cellular respiration in M. marinum, we created a transgenic M. marinum Δesx1 strain carrying pMV762Peredox-mCherry (Addgene 90217) to visualise build up of NADH:NAD ratio [34,35].

Techniques: Gene Expression, Infection, Expressing, Mutagenesis

Journal: Free radical biology & medicine

Article Title: The cyclic nitroxide antioxidant 4-methoxy-TEMPO decreases mycobacterial burden in vivo through host and bacterial targets.

doi: 10.1016/j.freeradbiomed.2019.03.010

Figure Lengend Snippet: Fig. 6. MetT has antibacterial effects against stressed bacteria. (A) CFU recovery from axenic early log phase cultures of M. marinum treated with MetT for 2 days. (B) Quantification of constitutive mCherry fluorescence in late log phase cultures of M. marinum Δesx1 strain carrying pMV762-Peredox-mCherry treated with MetT. (C) Quantification of peredox fluorescence in late log phase cultures of M. marinum Δesx1 strain carrying pMV762-Peredox-mCherry treated with MetT as an indicator of cellular NADH:NAD build up. (D) CFU recovery from early log phase cultures of M. marinum treated with MetT for 2 days. (E) Representative images of 4 dpi zebrafish embryos infected with M. marinum-wasabi and exposed to 10% O2 in a hypoxia chamber. (F) Quantification of bacterial burden by fluorescent pixel count in 4 dpi zebrafish embryos treated MetT and maintained in 10% O2 as indicated. Each dot represents one embryo.

Article Snippet: To visualise disruption of cellular respiration in M. marinum, we created a transgenic M. marinum Δesx1 strain carrying pMV762Peredox-mCherry (Addgene 90217) to visualise build up of NADH:NAD ratio [34,35].

Techniques: Bacteria, Infection