Journal: Cell death & disease

Article Title: Caspase-8 activation by cigarette smoke induces pro-inflammatory cell death of human macrophages exposed to lipopolysaccharide.

doi: 10.1038/s41419-023-06318-6

Figure Lengend Snippet: Fig. 2 hMDMs exposed to CSE and LPS display apoptotic hallmarks. Representative images of hMDMs treated with 1 μg/ml of LPS and 20% CSE, alone or in combination, for 24 h and stained with (A) Hoechst 33342 for nuclear morphology evaluation and (B) JC-1 dye for MMP assessment. C Quantification of Hoechst 33342 intensity expressed as mean fluorescent intensity (MFI). D Quantification of the red (485/ 590 nm)/green (485/529 nm) fluorescence intensity ratio after JC1 staining. Scale bar: 50 μm. Data are presented as mean ± SEM (N = 3 independent donors).

Article Snippet: Hoechst fluorescent nucleic acid stain (33342) was obtained from ImmunoChemistry technology, Davis, California, USA.

Techniques: Staining

Journal: Bioorganic & medicinal chemistry

Article Title: Advances in the structural studies of antibiotic potentiators against Escherichia coli

doi: 10.1016/j.bmc.2019.06.003

Figure Lengend Snippet: Assay for inhibition of the efflux of the fluorophore Hoechst 33342 (H33342) by 7 (A) or 22 (B) in E. coli ATCC 25922. All reactions were initiated by the addition of 50 mM glucose, except where indicated. H33342 was used at a concentration of 10 μM. Both compounds were used at concentrations ranging from 1/32X to ¼X MIC (1.56 to 12.5 μg/ml for 7 and 3.125 to 25 μg/ml for 22). The intrinsic MICs of 7 and 22 against E. coli ATCC 25922 are 50 and 100 μg/ml, respectively. Compound vehicle was DMSO.

Article Snippet: Examination of the impact that increasing concentrations of 7 and 22 have on the fluorescence decrease stimulated by the addition of glucose reveals that both compounds inhibit pump-mediated efflux of {"type":"entrez-nucleotide","attrs":{"text":"H33342","term_id":"978759","term_text":"H33342"}} H33342 to a similar degree, with the extent of this inhibition being dependent on the concentration of the test compound ( ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 2. caption a7 Assay for inhibition of the efflux of the fluorophore Hoechst 33342 ( {"type":"entrez-nucleotide","attrs":{"text":"H33342","term_id":"978759","term_text":"H33342"}} H33342 ) by 7 (A) or 22 (B) in E. coli ATCC 25922.

Techniques: Inhibition, Concentration Assay

Journal: Reproduction (Cambridge, England)

Article Title: mTORC1-dependent suppression of autophagic activity in somatic cell nuclear transfer mouse embryos

doi: 10.1530/REP-25-0338

Figure Lengend Snippet: Autophagy dynamics at the 2-cell to 4-cell stage in IVF and SCNT embryos. (A) Schematic illustration of the experimental process. IVF (top) and SCNT (bottom) were used to fix early 2-cell, late 2-cell, and 4-cell samples after 18, 32, and 42 h, respectively, with 0 h of insemination or activation. (B) IVF and SCNT embryos at the 2-cell to 4-cell stage co-stained for LC3 (green) and LAMP1 (magenta). White arrowheads were autophagosomes, black arrowheads were autolysosomes, and DNA was stained with Hoechst 33342. The magnified image shows yellow squares in the bottom panels. (C) Number of LC3 puncta at the 2-cell to 4-cell stage. At least ten embryos per developmental stage were used. Significant differences were determined by a two-tailed t -test. * P < 0.05. Error bars indicate SEM. (D) mRNA expression profiles of Map1lc3b and Lamp1 in preimplantation fertilized embryos. RNA-seq data were from publicly available sources ( Deng et al. 2014 ).

Article Snippet: The nuclei were co-stained with Hoechst 33342 (H342, Dojindo, Japan).

Techniques: Activation Assay, Staining, Two Tailed Test, Expressing, RNA Sequencing

Journal: Reproduction (Cambridge, England)

Article Title: mTORC1-dependent suppression of autophagic activity in somatic cell nuclear transfer mouse embryos

doi: 10.1530/REP-25-0338

Figure Lengend Snippet: Expression of TORC1 signaling genes and intracellular dynamics of mTOR in IVF and SCNT embryos. (A) Line plots showing gene set enrichment analysis (GSEA) of TORC1 signaling (GO:0038202) for SCNT and IVF embryos. The upper column shows the 2-cell stage, and the lower column shows the 4-cell stage, with normalized enrichment score (NES) and FDR q -value displayed, respectively. (B) Heatmap showing gene expression at the 4-cell stage of IVF and SCNT embryos, and cumulus cells used as donor cells for SCNT, for the gene groups analyzed in A. TPM values were converted to Z-scores for each gene. The K-means method was used to divide the data into four clusters. (C) Immunostaining for mTOR (green) and the lysosomal marker LAMP1 (magenta) in IVF and SCNT embryos at the 2-cell to 4-cell stage. DNA was stained with Hoechst 33342. The magnified image shows yellow squares in the 4-cell stage. (D) Line profile showing fluorescence intensity along the yellow line drawn in the magnified image in C. The fluorescence intensity of mTOR (green) and LAMP1 (magenta) is shown at 100 pixels from the yellow point to the white point, respectively.

Article Snippet: The nuclei were co-stained with Hoechst 33342 (H342, Dojindo, Japan).

Techniques: Expressing, Gene Expression, Immunostaining, Marker, Staining, Fluorescence