Journal: Nature Communications

Article Title: Creation of a biological sensorimotor interface for bionic reconstruction

doi: 10.1038/s41467-024-49580-8

Figure Lengend Snippet: A cross-section of a nerve fiber bundle in the dermal graft. It contains many small- to medium-sized NF-positive nerve fibers (red), however, only some have a myelin sheath as visualized by the anti-MBP staining (green). ( n = 12, biological replicates) B Two reinnervated muscle spindles (large arrows) within their respective capsules. They are accompanied by bundles of large, myelinated (S100, green) nerve fibers (NF, red) (*). These types of fibers can also be seen attached to the intrafusal fibers (small arrows), suggesting robust proprioceptive reinnervation. ( n = 13, biological replicates) (NF neurofilament, MBP myelin basic protein).

Article Snippet: Mechanoreceptors and muscle spindles were stained using 10% goat serum (Dako, Agilent Technologies, USA) in combination with a Dako Omnis rabbit anti-S100 primary antibody (Dako, Agilent Technologies, USA) solution (1:1 in PBST) or a mouse anti-MBP primary antibody (Santa Cruz Biotechnology, USA) solution (1:500 in PBST), respectively, in addition to the aforementioned anti-NF primary antibody.

Techniques: Staining, Capsules

Journal: STAR Protocols

Article Title: Protocol for detecting histidine polyphosphate modification of human proteins via MBP-tagged expression in E. coli

doi: 10.1016/j.xpro.2024.102947

Figure Lengend Snippet:

Article Snippet: Incubate membrane with primary antibody, mouse anti-MBP (1/1,000) (Cell Signaling, Cat#2396S) rocking for 18 h at 4°C. iii.

Techniques: Virus, Recombinant, Purification, Plasmid Preparation, Chromatography, Staining

Journal: Frontiers in Genetics

Article Title: The Pathogenic R3052W BRCA2 Variant Disrupts Homology-Directed Repair by Failing to Localize to the Nucleus

doi: 10.3389/fgene.2022.884210

Figure Lengend Snippet: The R3052W protein localizes to the cytosol. (A) Immunofluorescent localization of BRCA2 in untreated BRCA2 knockout cells (BRCA2 −/− ), and stable cell lines expressing either BRCA2 WT or R3052W. Representative images of 2XMBP-BRCA2 (red, anti-MBP), RAD51 (green), and nuclei (blue). (B) Live images of BRCA2 knockout cells expressing either BRCA2 WT or R3052W fused to GFP at the N-terminus. (C) Representative immunofluorescence images of laser micro-irradiation experiments in BRCA2 knockout cells stably expressing BRCA2 WT or R3052W. DNA damage (stripes) are depicted in red (gammaH2AX), green (RAD51), and nuclei in blue (DAPI). (D) Quantification of RAD51 fluorescence intensity in damage areas (stripes) over the background in non-irradiated areas of respective laser micro-irradiated nuclei. Each data point represents a single analyzed nucleus, while the solid line is a mean value ± SD (Kruskal-Wallis test with Dunn’s multiple comparison post hoc test; **** p value < 0.0001). (E) Quantification of RAD51 intensity in the nuclear and cytoplasmic compartments. Each data point represents a single analyzed area (220.16 microns × 220.16 microns), while bars represent mean ± SD (one-way ANOVA with Holm-Šídák’s multiple comparisons post hoc test, * p -value<0.05; ** p -value<0.01).

Article Snippet: Washes and antibody incubations were done with 1X TBS-T. Primary mouse antibodies against MBP (NEB E8032L, 1:5,000), RAD51 (Novus Biologicals 14b4, 1:1,000), and primary rabbit antibody against BRCA2 (Abcam, ab27976), GAPDH (Sta Cruz Biotechnology 0411m #sc-47724, 1:1,000) and HA (Cell Signaling C2974 mAB#3724, 1:1,000) were used for western blotting.

Techniques: Knock-Out, Stable Transfection, Expressing, Immunofluorescence, Irradiation, Fluorescence

Journal: Frontiers in Genetics

Article Title: The Pathogenic R3052W BRCA2 Variant Disrupts Homology-Directed Repair by Failing to Localize to the Nucleus

doi: 10.3389/fgene.2022.884210

Figure Lengend Snippet: R3052W cytoplasmic localization is not altered by CRM1 depletion or leptomycin treatment and retains binding to DSS1. (A) Immunofluorescent localization of BRCA2 in untreated stable cell lines expressing WT, D2723H, or R3052W BRCA2 proteins upon RNA interference-mediated depletion of CRM1 or treatment with the nuclear export inhibitor leptomycin B. Representative images of BRCA2 (red, MBP antibody) and DAPI staining to visualize nuclei (blue). (B) Western blots of total cellular lysates (TCL) and amylose pulldowns from HEK 293T cells transiently transfected with 2XMBP-BRCA2 WT, D2723H, or R3052W co-transfected with HA-DSS1. Anti-MBP antibody was used for BRCA2 detection, Anti-RAD51 antibody was used for endogenous RAD51 detection and Anti-HA antibody was used for DSS1 detection.

Article Snippet: Washes and antibody incubations were done with 1X TBS-T. Primary mouse antibodies against MBP (NEB E8032L, 1:5,000), RAD51 (Novus Biologicals 14b4, 1:1,000), and primary rabbit antibody against BRCA2 (Abcam, ab27976), GAPDH (Sta Cruz Biotechnology 0411m #sc-47724, 1:1,000) and HA (Cell Signaling C2974 mAB#3724, 1:1,000) were used for western blotting.

Techniques: Binding Assay, Stable Transfection, Expressing, Staining, Western Blot, Transfection