Journal: ACS Biomaterials Science & Engineering

Article Title: Potential Use of Exosomes as Diagnostic Biomarkers and in Targeted Drug Delivery: Progress in Clinical and Preclinical Applications

doi: 10.1021/acsbiomaterials.1c00217

Figure Lengend Snippet: Exosome drug-loading techniques. (A) Exosome-secreting cells or exosomes incubated with the desired cargos. Cargos diffuse across the cell and exosomal membrane and are subsequently packaged within the exosomes. (B) Desired nucleic acids can be loaded into exosomes via a transfection-based strategy. Transfected with vectors, the donor cell generates RNAs/proteins and packages these products into exosomes using endogenous expression and sorting machinery of the donor cell, respectively. Exosomes can be directly transfected with small RNAs for cargo loading purposes. (C) Cargos can be loaded into exosomes directly through physical treatments. Electroporation, sonication, and surfactant treatment generate pores on the exosomal membranes that facilitate cargo loading. Freeze–thaw treatment, extrusion, and dialysis enhance cargo loading into exosomes during membrane recombination processes. Adapted from Fu et al. ref . Copyright 2020 Elsevier.

Article Snippet: The electroporation buffer can be trehalose pulse medium (TPM; 50 mM trehalose (Sigma-Aldrich, Cat. No. T0167) in PBS) or (1.15 mM potassium phosphate, pH = 7.2, 25 mM potassium chloride, 21% Optiprep) or cytomix electroporation buffer (120 mM KCl, 0.15 mM CaCl 2 , 10 mM KPO 4 , 25 mM HEPES, 2 mM EGTA, and 5 mM MgCl2, adjusted to pH 7.6 with KOH)., Then electroporation is carried out using a GenePulser Xcell electroporator (e.g., from Bio-Rad).

Techniques: Incubation, Membrane, Transfection, Expressing, Electroporation, Sonication

Journal: Journal of Biological Chemistry

Article Title: XRCC2 Is a Nuclear RAD51-like Protein Required for Damage-dependent RAD51 Focus Formation without the Need for ATP Binding

doi: 10.1074/jbc.m102396200

Figure Lengend Snippet: FIG. 2. Damage-dependent RAD51 focus formation. A, foci in wild-type V79, irs1, and irs1 cells transfected with XRCC2 with and without x-ray exposure (10 Gy/5 h). Representative fields are shown. B, time course of focus formation following 10-Gy x-rays. Results from representative experiments are shown. PAC, P1-artificial chromosome.

Article Snippet: Approximately 4 mg of plasmid DNA was transfected into irs1 cells by electroporation (Bio-Rad Gene Pulser at 400 V/500 microfarads) in Cytomix buffer (20).

Techniques: Transfection

Journal: Journal of Biological Chemistry

Article Title: XRCC2 Is a Nuclear RAD51-like Protein Required for Damage-dependent RAD51 Focus Formation without the Need for ATP Binding

doi: 10.1074/jbc.m102396200

Figure Lengend Snippet: FIG. 3. A, cell cycle distributions of V79 and irs1 cells in response to 10-Gy x-rays. B, Western blot of RAD51 in V79, irs 1, and irs1 cells transfected with XRCC2 with and without 10-Gy irradiation. In a representative experiment using the a-tubulin response as a loading control, the RAD51 signal strength showed no significant differences between cell lines or before and after irradiation.

Article Snippet: Approximately 4 mg of plasmid DNA was transfected into irs1 cells by electroporation (Bio-Rad Gene Pulser at 400 V/500 microfarads) in Cytomix buffer (20).

Techniques: Western Blot, Transfection, Irradiation, Control

Journal: Journal of Biological Chemistry

Article Title: XRCC2 Is a Nuclear RAD51-like Protein Required for Damage-dependent RAD51 Focus Formation without the Need for ATP Binding

doi: 10.1074/jbc.m102396200

Figure Lengend Snippet: FIG. 5. Survival curves for V79, irs1, and irs1 cells transfected with different wild-type and mutant XRCC2 cDNAs. A, mitomy- cin-C (mean and range of 2–3 experiments). B, x-rays (single experi- ments). wt, wild type.

Article Snippet: Approximately 4 mg of plasmid DNA was transfected into irs1 cells by electroporation (Bio-Rad Gene Pulser at 400 V/500 microfarads) in Cytomix buffer (20).

Techniques: Transfection, Mutagenesis

Journal: Journal of Biological Chemistry

Article Title: XRCC2 Is a Nuclear RAD51-like Protein Required for Damage-dependent RAD51 Focus Formation without the Need for ATP Binding

doi: 10.1074/jbc.m102396200

Figure Lengend Snippet: FIG. 6. RAD51 focus formation in V79, irs1, and irs1 cells trans- fected with XRCC2 cDNA (wild-type or mutant forms). Means and standard error of data from 2–5 experiments for unirradiated and irradiated (10 Gy/5 h) cells.

Article Snippet: Approximately 4 mg of plasmid DNA was transfected into irs1 cells by electroporation (Bio-Rad Gene Pulser at 400 V/500 microfarads) in Cytomix buffer (20).

Techniques: Mutagenesis, Irradiation