Journal: PLOS Pathogens

Article Title: Hepatitis B virus hijacks MRE11–RAD50–NBS1 complex to form its minichromosome

doi: 10.1371/journal.ppat.1012824

Figure Lengend Snippet: Antibodies used for western blotting.

Article Snippet: Anti-NBS1 Mouse Monoclonal antibody , Proteintech, Wuhan, China , Cat# 66980-1-Ig.

Techniques: Western Blot

Journal: PLOS Pathogens

Article Title: Hepatitis B virus hijacks MRE11–RAD50–NBS1 complex to form its minichromosome

doi: 10.1371/journal.ppat.1012824

Figure Lengend Snippet: (A) Schematic diagram of biotinylated HBV rcDNA pull-down and mass spectrometry assay. (B) GO analysis of target proteins were performed using Metascape. Bar graph of enriched terms across input gene lists, colored by p-values. (C) Protein-protein interaction network and molecular complex detection (MCODE) components identified in the gene lists. The three components of MRN complex: MRE11, RAD50 and NBS1 were labeled in red. The interaction between HBV rcDNA and MRN complex was further confirmed by western blotting (D) and chromatin immunoprecipitation assay (E), the numbers below the blot indicate the arbitrary units from the densitometry analysis of indicated bands. ***: p <0.001.

Article Snippet: Anti-NBS1 Mouse Monoclonal antibody , Proteintech, Wuhan, China , Cat# 66980-1-Ig.

Techniques: Mass Spectrometry, Labeling, Western Blot, Chromatin Immunoprecipitation

Journal: Theranostics

Article Title: Single-cell RNA-Seq analysis of molecular changes during radiation-induced skin injury: the involvement of Nur77

doi: 10.7150/thno.100417

Figure Lengend Snippet: Changes in the transcriptional profiles of skin fibroblast during radiation-induced skin injury. (A) Bubble plot of TFs alterations in rat and human skin cells with or without radiation. (B) Venn plots showing the number of shared upregulated (upper) and downregulated (lower) DEGs between different groups of human and rat skin samples. Violin plots showing the expression levels of 5 common DEGs in human and rat skin. (C) The t-SNE plot displays rat (left) and human (right) fibroblast. Bar plots showing the cell number of each cell subtypes contributed. (D) Violin plot showing Nur77 gene expression changes across different fibroblast subcluster in rat (left) and human (right). (E) Pseudotime ordering on rat fibroblasts and human fibroblasts (F) arranged them into a major trajectory, with two minor bifurcations. Each dot represents a single cell. The black arrow indicates the start and direction of the trajectory. Feature plots of expression distribution for Nur77 across pseudotime. (G) Heatmap showing the top 5 markers for fibroblast subcluster from the rat (left) and human (right) skin.

Article Snippet: Cells were blocked with blocking buffer (phosphate buffered saline, 1% Triton X-100, and 5% BSA) and incubated at 4 °C with Nur77 (#NBP2-66980, Novus) antibody (1: 200) overnight, FITC-conjugated goat anti-mouse antibody (1:300) was added for 2 h at room temperature, and cells were dyed with 200 nM MitoTracker Red CMXRos (#C1035, Beyotime) at 37 °C for 30 min in darkness.

Techniques: Expressing

Journal: Theranostics

Article Title: Single-cell RNA-Seq analysis of molecular changes during radiation-induced skin injury: the involvement of Nur77

doi: 10.7150/thno.100417

Figure Lengend Snippet: Nur77 is involved in the irradiation process of skin cells. (A) qRT-PCR analysis of Nur77 mRNA expression in response to radiation in WS1 and HaCaT cells. (B) Western blotting analysis showing Nur77 expression in WS1 and HaCaT cells after different time post irradiation or different dose of irradiation. (C) and (D) Detection of radiation-induced nuclear-cytoplasmic distribution of Nur77 determined by preforming immunofluorescence analysis separating nuclear and cytoplasmic fraction and separating nuclear and cytoplasmic fractions. (E) Expression of Nur77 in normal and irradiated human skin tissues. (F) Western blot analysis showing the distribution of Nur77 in different organs and changes over time from 3 to 6 days after irradiation. (G) The effect of C-DIM8 on ROS production after different dose of irradiation as determined by DCFH-DA staining in WS1 and HaCaT cells. (H) The effect of C-DIM8 on cell apoptosis determined by AV/PI staining in WS1 and HaCaT cells. (I) The effect of Nur77 inhibitor C-DIM8 on radiosensitivity as determined by a colony formation assay following different doses of radiation. (J) Western blotting analysis showing cell death-related biomarker expression in irradiated WS1 cells treated with C-DIM8. * P < 0.05 and ** P < 0.01, compared with the control group. Scale bar = 200 μm.

Article Snippet: Cells were blocked with blocking buffer (phosphate buffered saline, 1% Triton X-100, and 5% BSA) and incubated at 4 °C with Nur77 (#NBP2-66980, Novus) antibody (1: 200) overnight, FITC-conjugated goat anti-mouse antibody (1:300) was added for 2 h at room temperature, and cells were dyed with 200 nM MitoTracker Red CMXRos (#C1035, Beyotime) at 37 °C for 30 min in darkness.

Techniques: Irradiation, Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Staining, Colony Assay, Biomarker Assay, Control

Journal: Theranostics

Article Title: Single-cell RNA-Seq analysis of molecular changes during radiation-induced skin injury: the involvement of Nur77

doi: 10.7150/thno.100417

Figure Lengend Snippet: Loss of Nur77 aggravates radiation-induced skin injury in mouse models. (A) The phenotypes and genotypes of wild-type ( Nur77 +/+ ) and Nur77 knockout ( Nur77 -/- ) mice. (B) Schematic of the workflow showing the establishment of the three mouse models with radiation-induced skin injury. 1) Acute radiation-induced skin injury; 2) Radiation fractionation; 3) A mouse model of full-thickness skin wounds combined with 4 Gy total-body irradiation. Loss of Nur77 aggravates radiation-induced skin injury in three mouse models. (C) Pictures showing the weight change and the scoring curves of the whole course of radiogenic injury in mice with different Nur77 genotypes. (D) Wound healing, body weight, and wound score results of radiation fractionation model in wild-type and Nur77 knockout mice. (E) Wound healing, body weight, and wound healing score results of the radiation combined injury model in mice. * P < 0.05 and ** P < 0.01, compared with the control group.

Article Snippet: Cells were blocked with blocking buffer (phosphate buffered saline, 1% Triton X-100, and 5% BSA) and incubated at 4 °C with Nur77 (#NBP2-66980, Novus) antibody (1: 200) overnight, FITC-conjugated goat anti-mouse antibody (1:300) was added for 2 h at room temperature, and cells were dyed with 200 nM MitoTracker Red CMXRos (#C1035, Beyotime) at 37 °C for 30 min in darkness.

Techniques: Knock-Out, Fractionation, Irradiation, Control

Journal: Theranostics

Article Title: Single-cell RNA-Seq analysis of molecular changes during radiation-induced skin injury: the involvement of Nur77

doi: 10.7150/thno.100417

Figure Lengend Snippet: scRNA-Seq reveals the complex mechanism by which Nur77 mediates radiation-induced skin injury. (A) Diagram displaying the process of sequencing single cells from radiation-induced skin injury samples obtained from wild-type ( Nur77 +/+ ) and Nur77 knockout ( Nur77 -/- ) mice. (B) The t-SNE plot displays main cell types in wild-type and Nur77 knockout mice. Each dot represents only one cell. (C) Dot plot showing the expression of representative genes for each cell type. (D) The U-MAP plot displays cell types mouse skin with or without radiation. Each dot represents only one cell. (E) Bar plots show the proportions that each group contributes to each cluster. (F) The Venn diagram shows the number of up-regulated DEpcGs and down-regulated DEpcGs in different cell types. (G) Significant signaling pathways were ranked based on differences in the overall information flow within the inferred networks between Nur77 -/- and Nur77 +/+ mouse skin. The overall information flow of a signaling network is calculated by summarizing all communication probabilities in that network. An overview of cell-cell interactions. Arrow and edge color indicate direction. Bar plots showing overall information flow of each signaling pathway. (H) Heatmap shows outgoing signaling patterns of Nur77 -/- and Nur77 +/+ mouse skin. (I) Comparison of the significant ligand-receptor pairs between Nur77 -/- and Nur77 +/+ mouse skin, which contribute to the signaling from fibroblast to other cells.

Article Snippet: Cells were blocked with blocking buffer (phosphate buffered saline, 1% Triton X-100, and 5% BSA) and incubated at 4 °C with Nur77 (#NBP2-66980, Novus) antibody (1: 200) overnight, FITC-conjugated goat anti-mouse antibody (1:300) was added for 2 h at room temperature, and cells were dyed with 200 nM MitoTracker Red CMXRos (#C1035, Beyotime) at 37 °C for 30 min in darkness.

Techniques: Sequencing, Knock-Out, Expressing, Comparison