Journal: Eukaryotic Cell

Article Title: Regulation of Cryptococcus neoformans Capsule Size Is Mediated at the Polymer Level

doi: 10.1128/ec.00437-07

Figure Lengend Snippet: FIG. 1. Electromobility of shed GXM from JEC21 cells changes over time in rich medium. All samples were heated (15 min at 70°C) to denature enzymes, centrifuged (16,000 g for 3 min) to separate supernatant and cells, and stored at 4°C. Samples (15 l) of culture supernatants were mixed with 6 DNA loading dye (4), loaded on a 0.6% certified megabase agarose (Bio-Rad) gel, and subjected to elec- trophoresis (15 h at 25 V) in 0.5 TBE (44.5 mM Tris base, 44.5 mM boric acid, 1 mM EDTA, pH 8.3). Samples containing more GXM (later time points) were diluted in distilled water based on a pilot gel, as sample normalization based on enzyme-linked immunosorbent as- say determination of GXM concentration (23) did not yield equal blot intensity. (This may reflect changes over time in GXM antibody reac- tivity [7, 11, 18] and/or transfer efficiency.) The gel was transferred onto a positively charged nylon membrane and immunoblotted with 1 g/ml anti-GXM antibody 3C2 as described in the text. Initial sample heating and dilution in distilled water did not alter electrophoretic migration (data not shown). St, starter culture (at 1.6 107 cells/ml); arrowhead, well position; arrow, direction of migration; other num- bers, sampling time (hours) after the start of the experiment.

Article Snippet: Samples (15 l) of culture supernatants were mixed with 6 DNA loading dye (4), loaded on a 0.6% certified megabase agarose (Bio-Rad) gel, and subjected to electrophoresis (15 h at 25 V) in 0.5 TBE (44.5 mM Tris base, 44.5 mM boric acid, 1 mM EDTA, pH 8.3).

Techniques: Concentration Assay, Membrane, Migration, Sampling

Journal: Cell

Article Title: Microglia Require CD4 T Cells to Complete the Fetal-to-Adult Transition

doi: 10.1016/j.cell.2020.06.026

Figure Lengend Snippet:

Article Snippet: 96.96 DNA Binding Dye Sample/Loading Kit—10 IFCs, with Control Line Fluid , Fluidigm , Cat#BMK-M10- 96.96-EG.

Techniques: Labeling, Plasmid Preparation, Purification, Recombinant, Binding Assay, Cell Isolation, Conjugation Assay, Sequencing, Transgenic Assay, Knock-Out, Software

Journal: Nature Communications

Article Title: Investigation of artificial cells containing the Par system for bacterial plasmid segregation and inheritance mimicry

doi: 10.1038/s41467-024-49412-9

Figure Lengend Snippet: a Giant unilamellar vesicles (GUV) containing the ParMRC system and PURE system. b parC -eGFP DNA segregation by the polymerization of ParM triggered by adenosine triphosphate (ATP) influx upon laser irradiation. c A deformed GUV under hypertonic conditions. d Two daughter GUVs containing parC -eGFP DNA and the PURE system upon laser irradiation at the center region of the deformed GUV (purple area in Fig. 1c). e Enhanced green fluorescent protein (eGFP) was expressed inside two daughter GUVs through translating the eGFP gene using a PURE (protein synthesis using recombinant elements) system at 37 °C. The PURE system contains ribosomes, amino acids, nucleoside triphosphates (NTPs), transfer ribonucleic acid (tRNAs), enzyme substrates, RNA polymerase, translation factors, and other necessary components.

Article Snippet: A Bradford protein assay kit, SDS‒PAGE gel configuration kit, 4×SDS‒PAGE sample loading buffer, protease inhibitor cocktail for purification of His-tagged proteins, green fluorescent DNA marker dye, streptavidin magnetic beads, Tris-borate-EDTA buffer (TBE) and Tween-20 were purchased from Beyotime (China).

Techniques: Irradiation, Recombinant

Journal: Nature Communications

Article Title: Investigation of artificial cells containing the Par system for bacterial plasmid segregation and inheritance mimicry

doi: 10.1038/s41467-024-49412-9

Figure Lengend Snippet: a Schematic illustration of two parC -eGFP DNAs being pushed through ParM polymerization in a giant unilamellar vesicles (GUV) and subsequent GUV division under laser irradiation. b Confocal microscopy images of GUV division ( b1 – b3 ) and enhanced green fluorescent protein (eGFP) expression at 37 °C ( b4 – b7 ). b1 – b3 indicate GUV deformation, filament splitting, and division into two daughter cells, respectively. The white arrows in b1 and b2 indicate parC -eGFP DNA. The ParM filament was split by laser irradiation (561 nm, 0.7 mW, 5 s). The scale bars are 10 μm. n = 3 independent replicates. c Schematic illustration of eGFP expression in two daughter cells. The corresponding fluorescence intensity (FI) in daughter GUV 1 ( d ) and daughter GUV 2 ( e ) as a function of time. The normalized fluorescence intensity of eGFP was calculated from three independent samples. The data were presented as the mean values ± SDs; n = 3 independent replicates. Source data are provided as a Source Data file.

Article Snippet: A Bradford protein assay kit, SDS‒PAGE gel configuration kit, 4×SDS‒PAGE sample loading buffer, protease inhibitor cocktail for purification of His-tagged proteins, green fluorescent DNA marker dye, streptavidin magnetic beads, Tris-borate-EDTA buffer (TBE) and Tween-20 were purchased from Beyotime (China).

Techniques: Irradiation, Confocal Microscopy, Expressing, Fluorescence