Journal:

Article Title: Alteration of Large-Scale Chromatin Structure by Estrogen Receptor

doi: 10.1128/MCB.22.10.3437-3449.2002

Figure Lengend Snippet: Histone acetylation and brahma recruitment by GFP-lac rep-ER fusion proteins. A03_1 cells were transfected with GFP-lac rep-ER, GFP-lac rep, or GFP-lac rep-VP16 and treated with 10−9 M estradiol for various time points prior to fixation and staining with antibodies against the acetylated forms of histone H3 and H4 or against brahma. (A) For histone acetylation (left set of images), each cell's lac operator array was scored as having staining at a lower level than (hypoacetylated), comparable to (neutral), or higher than (hyperacetylated) that for the rest of the chromatin in the nucleus. For brahma recruitment (right set of panels), each cell's lac operator array was scored as having staining comparable to that for the rest of the nucleus (not recruited), slightly brighter staining relative to the rest of the nucleus (borderline), or brighter staining than the rest of the nucleus (recruited). (B) For each time point, cells were scored as in panel A for histone H3 acetylation (left) and histone H4 acetylation (middle) and brahma recruitment (right).

Article Snippet: Coverslips were washed three times (5 min each) in 0.1% Triton X-100 and incubated overnight at 4°C with a primary antibody diluted in 0.1% Triton X-100 as follows: 1:500 antiacetylated histone H3 (AHP412; Serotec, Raleigh, N.C.), 1:500 antiacetylated histone H4 (AHP418; Serotec), 1:50 anti-brahma N-19 (sc-6450; Santa Cruz Biotechnology, Santa Cruz, Calif.), 1:100 anti-BAF170 C-19 (sc-9744; Santa Cruz Biotechnology).

Techniques: Transfection, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Streptococcus suis Serotype 2 Biofilms Inhibit the Formation of Neutrophil Extracellular Traps

doi: 10.3389/fcimb.2017.00086

Figure Lengend Snippet: NETs visualization and quantification. (A) Neutrophils were stimulated with planktonic SS2. The pictures from the left to right were labeled with the following dyes: DNA with DAPI (blue), histone H4 (citrulline 3) with Alexa 568 conjugated (red), and an overlay of the first two pictures using ZEN 2012 software (Zeiss). Scale bar, 10 μm. (B) At 100 × magnification with oil, DNA was stained with the SYTO 9 green fluorescent nucleic stain. Arrows indicate the NETs structure; the round shapes indicate free bacteria without entrapment, and the square shapes indicate bacteria entrapped by NET DNA. (C) Relative fluorescence units were used to evaluate the quantity of NETs. Planktonic SS2 and bacteria separated from biofilm matrix could induce NETs release. The NET formation level induced by planktonic SS2 with PMA was twice that induced by planktonic SS2. The NET formation induced by bacteria separated from biofilm matrix with PMA-treated neutrophils was twice that induced by bacteria only. NETs induced by biofilm SS2 and the biofilm matrix were similar to the negative control. The results are depicted as the mean ± SD ( n = 5). ** p < 0.01; *** p < 0.001; ns, no difference between groups.

Article Snippet: The samples were stained with the primary rabbit anti-neutrophil histone H4 antibody (citrulline 3, 1:1000 diluted, Merck Millipore, Billerica, MA, USA) for 1 h at RT, followed by incubation with goat anti-rabbit Alexa 568 antibody (1:100 dilution, Jackson ImmunoResearch, West Grove, PA, USA).

Techniques: Labeling, Software, Staining, Bacteria, Fluorescence, Negative Control