Journal: bioRxiv

Article Title: Sialidases derived from Gardnerella vaginalis remodel the sperm glycocalyx and impair sperm function

doi: 10.1101/2025.02.01.636076

Figure Lengend Snippet: A) Schematic describing lectin flow cytometry and representative glycan epitopes. B, C, D) Sperm were exposed to sialidases (AUS 20 units or GvNanH2 160 units) for 1 hour, fixed in formalin, and stained with MAL-II-biotin/Neutravidin-Alexa Fluor 488, SNA-Cy5, or PSA-FITC respectively. Each point represents the median fluorescence intensity from one donor. Statistics represent a mixed model comparison with post-hoc Holm-Šidák tests. E) Schematic representing change in zeta potential after removal of negatively charged sialic acid on sperm surface. F) Sperm were treated with sialidase at various doses for 1 hour, and zeta potential was measured via dynamic light scattering. Each measurement represents the average of seven technical replicates, and statistics represent a mixed model comparison with post hoc Holm-Šidák tests.

Article Snippet: Biotinylated Maackia Amurensis Lectin II (MAL II, Cat #:B-1265-1) and Cy5-conjugated Sambucus Nigra Lectin (SNA, Cat #: CL-1305-1) were purchased from Vector Labs. Human Contraception Antibody (HCA), was a gift from ZabBio.

Techniques: Flow Cytometry, Staining, Fluorescence, Comparison, Zeta Potential Analyzer

Journal: PLoS ONE

Article Title: Transcriptomic Analysis of Liquid Non-Sporulating Streptomyces coelicolor Cultures Demonstrates the Existence of a Complex Differentiation Comparable to That Occurring in Solid Sporulating Cultures

doi: 10.1371/journal.pone.0086296

Figure Lengend Snippet: (A) Liquid non-sporulating cultures. (B) Solid sporulating cultures. Mycelial structures (MI, first compartmentalized mycelium; MII, second multinucleated mycelium). The classical nomenclature of substrate, aerial mycelium, and hydrophobic layers are indicated. Three independent biological replicates and two developmental stages (MI and MII) were processed and cDNA was labeled with Cy3 while chromosomal S. coelicolor DNA was used as reference and labeled with Cy5. The scheme was adapted from Manteca et al. .

Article Snippet: Genomic DNA from S. coelicolor M145 was used as the common reference. gDNA was labeled with Cy5-dCTP (GE Healthcare Life Sciences) using the BioPrime Array CGH Genomic Labeling Module (Invitrogen), purified with the MinElute kit, and labeling efficiencies quantified with a NanoDrop ND-1000 spectrophotometer.

Techniques: Labeling

Journal: Frontiers in molecular neuroscience

Article Title: Specific Role for GSK3α in Limiting Long-Term Potentiation in CA1 Pyramidal Neurons of Adult Mouse Hippocampus.

doi: 10.3389/fnmol.2022.852171

Figure Lengend Snippet: FIGURE 1 | Spatial validation of GSK3α or GSK3β cKO by IHF. (A) Fluorescence images of brain sections through the hippocampus collected with a 10 × objective lens following staining with anti-GSK3α (revealed with Alexa 594) and anti-GSK3β (revealed with Cy5), as well as DNA using DAPI; the three columns on each row is from a single section imaged sequentially in the corresponding fluorescence wavelength channels. IHF from each of a GSK3α cKO, GSK3β cKO, and a Cre- subject are shown in rows. Note the prominent reduction in corresponding GSK3 staining within the CA1 region of each paralog cKO. (B) The white outlined zones in A were subsequently imaged with a 60 × lens and confocal optical resolution. The same fluorescence channels are presented, plus a final merged overlay in the final column. (C) Primary antibody was omitted, and images from the same Cre- subject were collected at the exact same settings as those used in (A). (A–C) Images shown were selected from a sampling of n = 2–3 subjects per genotype, and are representative from a total of 36 images collected with a 10 × objective lens, and 82 images collected with a 60 × objective lens.

Article Snippet: Slides were washed with PBS 3 × 5 min, incubated with secondary antibodies, donkey anti-rabbit conjugated with Alexa Fluor 594 (Jackson ImmunoResearch Inc, West Grove, PA, United States, Cat# 711- 585-152) and donkey anti-mouse conjugated with Cy5 (Jackson ImmunoResearch Inc, Cat# 715-175-150) at 1:1000 diluted with PBS for 1 h at room temperature.

Techniques: Biomarker Discovery, Fluorescence, Staining, Sampling