Journal:

Article Title: Effect of notoginsenoside R1 on hepatic microcirculation disturbance induced by gut ischemia and reperfusion

doi: 10.3748/wjg.14.29

Figure Lengend Snippet: Effect of R1 on the expression of E-selectin in hepatic vessels (A) and CD18 and CD11b in neutrophils (B) of mice subjected to SMA I/R. The results are presented as mean ± SE from 6 animals. aP < 0.05 vs control, cP < 0.05 vs I/R.

Article Snippet: Other reagents used in experiments were as follows: rhodamine 6G (purity > 99.0%, Lot No.2350994, Fluka Co., Switzerland), FITC-rat anti-mouse CD18 monocolonal antibody (Lot No.553293, BD Biosciences PharMingen, USA), FITC-rat anti-mouse CD11b monoclonal antibody (Lot No.557396, BD Biosciences PharMingen, USA), goat polyclonal antibody against mouse E-selectin (M-20) (sc-6939, Santa Cruz Biotechnology, Inc. USA), goat polyclonal antibody against mouse ICAM-1 (M-19) (sc-1511, Santa Cruz Biotechnology, Inc. USA), rhodamine conjugated rabbit anti-goat lgG-R (Lot No.B1006, Santa Cruz Biotechnology, Inc. USA), Hoechst33342 (Lot No.6538, Santa Cruz Biotechnology, Inc. USA), mouse MCP-1 flex set (Lot No.558342, BD Biosciences, USA), mouse TNF flex set (Lot No.558299, BD Biosciences, USA), mouse IL-6 flex set (Lot No.558301, BD Biosciences, USA).

Techniques: Expressing

Journal: Neuroscience bulletin

Article Title: Inflammatory Factor IL1α Induces Aberrant Astrocyte Proliferation in Spinal Cord Injury Through the Grin2c/Ca 2+ /CaMK2b Pathway.

doi: 10.1007/s12264-023-01128-4

Figure Lengend Snippet: Fig. 8 Schematic showing that after spinal cord injury, IL1α induces astrocyte proliferation by inhibiting Grin2c, result- ing in decreased intracel- lular Ca2+ concentration and CaMK2b levels. In response to the blockade of IL1α via neutralizing antibodies, Grin2c/ Ca2+/CaMK2b pathway levels increase, and astrocyte prolif- eration decreases.

Article Snippet: In particular, mice were intraperitoneally injected with an anti-mouse IL1α neutralizing antibody (R&D systems, Minneapolis, MN, SA, AF-400-NA, 20 mg/kg) and IgG control (R&D systems, Minneapolis, MN, SA, AB-108-C, 20 mg/kg) once per day for one week from the 1st day after surgery.

Techniques: Concentration Assay

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: a Gating strategy in screening experiment using the panel of TCR Vβ antibodies. First, CD3 lymphocyte were separated into CD4 + and CD8 + populations using monoclonal antibodies (CD8 + : Pe-Cy7, CD4 + : PerCP). Next, the samples were stained with Vβ specific antibody cocktails included in IO Test Beta Mark TCR beta Repertoire Kit (Beckman-Coulter Immunotech, USA). The β variable chain family was determined based on FITC and PE positivity from CD4 + and CD8 + populations according to the manufacturer’s instruction. Vβ20 clone was detected from total CD4 + T cells (52.9%, middle panel) and total CD8 + T cells (1.74%, right). b Flow cytometry Vβ screening results from the index patient’s peripheral blood sample. T cell clonality with antibodies which target Vβ region of TCR was analysed of CD4 + T cells. The increased distribution suggests that the cells have large T cell clone. c Increased Vβ20 bearing clonotype over time in the index patient’s CD4 + T cells. Source data are provided as a Source data file. d T cell repertoire of FACS-sorted CD4 + Vβ20+ and CD8 + T cells analysed with TCRβ deep sequencing (Adaptive Biotechnologies). The TCRBV30-01 clone was detected in the CD4 + Vβ20+ fraction, but not in the CD8 + fraction. e Multicolor flow cytometry was applied to identify the immune phenotype of HSCT donor and index patient’s memory T cell subtypes. Central memory (CM), naïve, effector memory (EM), and terminal effector memory (TEMRA) cells. f The relative proportion of granzyme B positive (GrB + ) CD4 + T cells and GrB + CD8 + T cells in index patient. Index patient’s PBMCs were stained with anti-CD45, −CD3, −CD4, and −CD8 (surface markers), and then GrB stained after fixation and permeabilization. Stained cells were analyzed using FACSVerse.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Staining, Flow Cytometry, Sequencing

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: Somatic mutations discovered in CD4 + T cells in the index patient, detected from 2013 sample.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Mutagenesis

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: a Locations of mTOR, TLR2 , and NFκB2 somatic mutations. Linearized structure of MTOR, NFκB2 , and TLR2 presenting the location of somatic mutations. mTOR P2229R mutation is located in the kinase domain, NFκB2 P882Q in the C-terminus, and TLR2 W558L between LRR (Leucine-rich repeats) domain and transmembrane (TM) domain. b A heterozygous mTOR mutation (G to C, P2229R ) was detected in CD4 + T cells by Sanger sequencing. c Variant allele frequencies (VAFs) of mTOR P2229R , NFκB2 P882Q , and TLR2 W558L mutations in the index patient’s CD4 + T cells over time as measured with amplicon sequencing. Source data are provided as a Source data file. d VAFs (%) of mTOR P2229R mutation from the index patient’s skin, liver, and eyes biopsy. e Immunofluorescence staining indicated CD3 + CD4 + and CD3 + CD8 + T cell infiltration in the skin. Paraffin embedded skin biopsy from the index patient was sectioned and stained with antibody specific human CD3 (cyan), CD4 (green), and CD8 (red). White arrows indicate infiltrated CD3 + CD4 + or CD3 + CD8 + T cells. Original imaging magnification: 20×, the figure has been further zoomed to 25× for visualization. Scale bar: 50 μm.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Mutagenesis, Sequencing, Variant Assay, Amplification, Immunofluorescence, Staining, Imaging

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: Somatic MTOR and NFκB2 mutations validated by amplicon sequencing in the index patient.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Amplification, Sequencing

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: Real-time cell analysing (RTCA) systems, xCELLigence TM , was applied to monitor real-time killing effect of primary fibroblasts obtained from the index patient and healthy donors. a Index patient’s primary fibroblasts were cultured as monolayers for 24 h to reach full confluence. Once confluent, the effector cells; NK-92 cell (positive control, 8:1), primary CD4 + T cells (4, 8, 16:1) and primary CD8 + T cells (16:1) with different ratios (effector cells:fibroblasts) were added to each well followed by co-culture (arrow indicates the point of adding the effector cells, 0 h). The control (black line, media) shows the impedance of the fibroblasts without any added effectors. The cell impedance was measured every 30 min for 21 h. The measured impedance was expressed as Cell Index with the normalization performed at time of addition of effector cells. b Visualization of monolayers of the primary fibroblast before and 21 h after addition of effector cells (8:1 for all effector cells). c CD4 + T cells and CD8 + T cells from healthy donors were added to healthy donors’ own fibroblasts ( n = 3). The cell impedance was measured for 21 h. Data is representative of three independent individuals. d Index patient’s fibroblasts were seeded with HLA1 and HLA2 antibodies (10 and 20 μg/mL). After CD4 + T cells (CD4 + T cells:fibroblasts = 8:1) were added the cell impedance was measured for 21 h. Dots represent mean values and error bars indicate range ( n = 2 for all conditions, technical duplicates). Source data are provided as a Source data file.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Cell Culture, Positive Control, Co-Culture Assay

Journal: Journal of Traditional Chinese Medicine

Article Title: Efficacy of Bushenjianpi prescription on autoimmune premature ovarian failure in mice

doi: 10.1016/S0254-6272(17)30321-7

Figure Lengend Snippet: Figure 3 Immunohistochemical analysis of the distinct presence of Cx43 and BMP-15 in the oophorons of each group (× 200) A-F: Cx43; G-L: BMP-15. A, G: control group; B, H: model group; C, I: positive group; D, J: low dose of BSJPP group; E, K: moderate dose of BSJPP group; F, L: high dose of BSJPP group. Positive group treated with premarin (0.03 mg/kg) once daily for 90 d. Con- trol and model group received the same volume of distilled water. Low (8.1 mg/kg), moderate (16.2 mg/kg, the dose used in vivo in human studies), and high (32.4 mg/kg) dose of BSJPP groups treated by gastrogavage once daily for 90 d. Immunohistochemi- cal analysis clearly revealed the presence of Cx43 and BMP-15 in the oophorons of control mice and those treated with BSJPP (D, E) and premarin compared with the model group. BSJPP: Bushenjianpi prescription; Cx43: connexin 43.

Article Snippet: Anti-connexin 43 (Cx43) antibody (product lot No. BA1727) and anti-bone morphogenetic protein 15 (BMP-15) antibody (product lot No. BA2018) were purchased from Boster Biological Engineering (Wuhan, China).

Techniques: Immunohistochemical staining, Control, In Vivo

Journal: Journal of Traditional Chinese Medicine

Article Title: Efficacy of Bushenjianpi prescription on autoimmune premature ovarian failure in mice

doi: 10.1016/S0254-6272(17)30321-7

Figure Lengend Snippet: Figure 4 Expression Cx43 and BMP-15 mRNA in the ovaries of mice in each group. 1: control group; 2: model group; 3: positive group; 4: low dose of BSJPP group; 5: moderate dose of BSJPP group; 6: high dose of BSJPP group. Positive group treated with pre- marin (0.03 mg/kg) once daily for 90 d. Control and model group received the same volume of distilled water. Low (8.1 mg/kg), moderate (16.2 mg/kg, the dose used in vivo in human studies), and high (32.4 mg/kg) dose of BSJPP groups treated by gastrogavage once daily for 90 d. Compared with the model group, significantly upregulated expression of both Cx43 and BMP-15 was detected in the control group and the groups treated with BSJPP (M and L groups). BSJPP: Bushenjianpi prescription; Cx43: connexin 43.

Article Snippet: Anti-connexin 43 (Cx43) antibody (product lot No. BA1727) and anti-bone morphogenetic protein 15 (BMP-15) antibody (product lot No. BA2018) were purchased from Boster Biological Engineering (Wuhan, China).

Techniques: Expressing, Control, In Vivo