Journal: The EMBO Journal

Article Title: A G1‐like state allows HIV ‐1 to bypass SAMHD 1 restriction in macrophages

doi: 10.15252/embj.201696025

Figure Lengend Snippet: A–D Peritoneal macrophages from 6‐ to 10‐week‐old C57BL/6 mice were stained for the macrophage markers (A) F4/80, (B) CD11b, (C) MCM2 or (D) active DNA synthesis (EdU) 2 h after isolation. Scale bars: 20 μm. E–K Peritoneal macrophages were isolated from 6‐ to 10‐week‐old WT or SAMHD1 −/− (KO) mice. Cells were infected with VSV‐G‐pseudotyped HIV‐1 GFP for 48 h and analysed for (G) infection, (H) MCM2 expression, (I) EdU incorporation (EdU was added at the time of infection), and co‐localisation between infection and (J) MCM2 protein or (K) active DNA synthesis (by EdU incorporation). (E, F) Random microscopic fields and plot profiles show an example of immunofluorescence intensity along infected cells. Scale bars: 10 μm. On average, 10 4 cells in each experiment were recorded and analysed using Hermes WiScan cell‐imaging system and ImageJ ( n ≥ 2, mean ± s.e.m.; (ns) non‐significant; **P ‐value ≤ 0.01, unpaired t ‐test).

Article Snippet: Antibodies used were as follows: anti‐cdc2 (Cell Signaling Technology, Beverly, MA, USA); anti‐CDK2 (H‐298, Santa Cruz Biotechnology); anti‐pCDK2(Thr160) (Bioss Inc., MA, USA); anti‐CDK4 (DCS156, Cell Signaling Technology); anti‐CDK6 (B‐10, Santa Cruz Biotechnology); anti‐SAMHD1 (ab67820, Abcam, UK), beta‐actin (ab6276, abcam, UK); mouse anti‐MCM2 (BM‐28, BD Biosciences, UK); and rabbit anti‐MCM2 (SP85) from Sigma; pSAMHD1 (a kind gift from M. Benkirane) and ProSci (Poway, CA, USA); anti‐Geminin (NCL‐L‐Geminin, Leica); anti‐mouse F4/80 and CD11b (kind gift from S. Yona, UCL); anti‐human CD68, CD14, CD163, CD80, CD86, CD40 (kind gift from M. Noursadeghi).

Techniques: Staining, DNA Synthesis, Isolation, Infection, Expressing, Immunofluorescence, Imaging

Journal: International journal of stem cells

Article Title: Integrin α 4 Positive Subpopulation in Adipose Derived Stem Cells Effectively Reduces Infarct Size through Enhanced Engraftment into Myocardial Infarction.

doi: 10.15283/ijsc22209

Figure Lengend Snippet: Fig. 2. Integrin α4 ＋ adipose-derived stem cells (ASCs) subpopulation were isolated and sorted from ASCs by fluo- rescence-activated cell sorting. (A) The population of integrin α4 ＋ ASCs were about 5.11% and 70.7% in the pre- and postsorted ASCs, respectively. (B) Immunofluorescent staining revealed integrin α4 were robustly expressed in integrin α4 ＋ ASCs subpopulation. (C) Western blotting showed that the ex- pression of integrin α4 with 135 kDa was obviously more in integrin α4 ＋

Article Snippet: A mouse anti-integrin α4 antibody was used to detect the grafted integrin α4+ ASCs. von Willebrand Factor (vWF), an endothelial cell marker, was identified by an mouse anti-vWF antibody (Proteintech).

Techniques: Derivative Assay, Isolation, FACS, Staining, Western Blot

Journal: International journal of stem cells

Article Title: Integrin α 4 Positive Subpopulation in Adipose Derived Stem Cells Effectively Reduces Infarct Size through Enhanced Engraftment into Myocardial Infarction.

doi: 10.15283/ijsc22209

Figure Lengend Snippet: Fig. 4. Identification of Integrin α4 ＋/green fluorescent protein＋ (GFP＋) adipose-derived stem cells (ASCs) and incorporation of injected ASCs into the vasculature by immunofluorescent staining in infarcted myocardium 4 weeks posttransplantation. (A) Immunofluorescent staining was performed for GFP (green) and integrin α4 (red), and nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI, blue), co-localized expression of integrin α4 and GFP were detected in the hearts receiving integrin α4 ＋ ASCs subpopulation, and integrin α4 ＋ ASCs sub- population showed much higher engraftment than unfractionated ASCs. (B) Immunofluorescent staining was conducted for GFP (green) and von Willebrand Factor (vWF, red), and nuclei were stained with DAPI (blue), GFP-positive ASCs directly incorporated into vasculature.

Article Snippet: A mouse anti-integrin α4 antibody was used to detect the grafted integrin α4+ ASCs. von Willebrand Factor (vWF), an endothelial cell marker, was identified by an mouse anti-vWF antibody (Proteintech).

Techniques: Derivative Assay, Injection, Staining, Expressing

Journal: International journal of stem cells

Article Title: Integrin α 4 Positive Subpopulation in Adipose Derived Stem Cells Effectively Reduces Infarct Size through Enhanced Engraftment into Myocardial Infarction.

doi: 10.15283/ijsc22209

Figure Lengend Snippet: Fig. 5. Integrin α4 ＋ adipose-derived stem cells (ASCs) subpopu- lation more robustly migrated and engrafted into the infartced my- ocardium after cell transplantation. (A) Four weeks posttranspla- ntation, green fluorescent protein-positive ASCs were more fre- quently detected in the hearts receiving integrin α4 ＋ ASCs sub- population than in the hearts undergoing unfractionated ASCs treatment. (B) The high reproducibility of the standard curve of re- al-time PCR. Serial dilution (10−2∼10−8) of DNA was made 7 times to construct the standard curve. Each pink circle corresponded to one dilution in one experiment. The blue solid line represented the regression analysis. (C) Four weeks after cell transplantation, in- tegrin α4 ＋ ASCs subpopulation treated rats showed the signifi- cantly greater ASC numbers per heart as compared to unfractio- nated ASCs treated rats. *p＜0.05 vs. unfractionated ASCs.

Article Snippet: A mouse anti-integrin α4 antibody was used to detect the grafted integrin α4+ ASCs. von Willebrand Factor (vWF), an endothelial cell marker, was identified by an mouse anti-vWF antibody (Proteintech).

Techniques: Derivative Assay, Transplantation Assay, Serial Dilution, Construct

Journal: International journal of stem cells

Article Title: Integrin α 4 Positive Subpopulation in Adipose Derived Stem Cells Effectively Reduces Infarct Size through Enhanced Engraftment into Myocardial Infarction.

doi: 10.15283/ijsc22209

Figure Lengend Snippet: Fig. 6. Integrin α4 + adipose-derived stem cells (ASCs) subpopulation more effectively reduced infarct size post-implantation in vivo. (A) Representative tomographic histograms of the 18F-fluorodeoxyglucose (FDG) positron emission tomography images at 4 weeks posttrans- plantation. Yellow colors were indicative of higher tracer uptake, while blue colors stood for lower tracer uptake. (B) Four weeks after transplantation, unfractionated ASCs transplantation markedly elevated the 18F-FDG uptake in apical-septal, apical-anterior, and apex seg- ments as compared with phosphate-buffered saline (PBS) control. Of importance, integrin α4 + ASCs subpopulation implantation further increased the 18F-FDG uptake in apical-septal, apical-inferior, and apex segments compared with unfractionated ASCs injection. (C) Representative transverse, coronal, and sagittal myocardial 18F-FDG images at 4 weeks after cell transplantation. The infarcted area can be visually detected as an area of low glucose metabolism. (D) Four weeks after transplantation, integrin α4 + ASCs subpopulation markedly reduced infarct size as compared to unfractionated ASCs and PBS control. (E) Integrin α4 + ASCs subpopulation significantly increased global 18F-FDG uptake compared with unfractionated ASCs and PBS control. *p＜0.05 vs. unfractionated ASCs. #p＜0.05 vs. PBS.

Article Snippet: A mouse anti-integrin α4 antibody was used to detect the grafted integrin α4+ ASCs. von Willebrand Factor (vWF), an endothelial cell marker, was identified by an mouse anti-vWF antibody (Proteintech).

Techniques: Derivative Assay, In Vivo, Positron Emission Tomography, Transplantation Assay, Saline, Control, Injection

Journal: International journal of stem cells

Article Title: Integrin α 4 Positive Subpopulation in Adipose Derived Stem Cells Effectively Reduces Infarct Size through Enhanced Engraftment into Myocardial Infarction.

doi: 10.15283/ijsc22209

Figure Lengend Snippet: Fig. 7. Integrin α4 ＋ adipose-derived stem cells (ASCs) subpopulation more effectively improve cardiac fucntion post-implantation in vivo. (A, B) Four weeks after transplantation, integrin α4 ＋ ASCs subpopulation reduced left ventricular end-diastolic volume (LVEDV) and left ventricular end-systolic volume (LVESV) compared with unfractionated ASCs and phosphate-buffered saline (PBS) control. (C) Integrin α4 ＋

Article Snippet: A mouse anti-integrin α4 antibody was used to detect the grafted integrin α4+ ASCs. von Willebrand Factor (vWF), an endothelial cell marker, was identified by an mouse anti-vWF antibody (Proteintech).

Techniques: Derivative Assay, In Vivo, Transplantation Assay, Saline, Control

Journal: eLife

Article Title: Near-perfect precise on-target editing of human hematopoietic stem and progenitor cells

doi: 10.7554/eLife.91288

Figure Lengend Snippet:

Article Snippet: Antibody , FITC anti-human, CD49c (Clone REA360) , Miltenyi , 130-105-364 , 1:50.

Techniques: Plasmid Preparation, Expressing, Sterility, Ligation, Selection, Sequencing, Recombinant, Electroporation, CRISPR, Software

Journal: General Psychiatry

Article Title: Microglia activation in the offspring of prenatal Poly I: C exposed rats: a PET imaging and immunohistochemistry study

doi: 10.1136/gpsych-2018-000006

Figure Lengend Snippet: (A) CD11b-immunopositive cells in the control group (A, C) and thetreatment group (B, D) Group comparison of OD values in the prefrontal cortex and hippocampus

Article Snippet: Then sections were incubated in a humidified chamber with the rabbit primary antibody against CD11b (1:100; Boster Biological Engineering, China) in PBS overnight at 4°C.

Techniques: Control, Comparison

Journal: Neural Regeneration Research

Article Title: Pre-clinical study of human umbilical cord mesenchymal stem cell transplantation for the treatment of traumatic brain injury: safety evaluation from immunogenic and oncogenic perspectives

doi: 10.4103/1673-5374.317985

Figure Lengend Snippet: Identification, immunogenicity and immunomodulation of huMSCs . (A) The huMSCs cultured in our experiment. The huMSCs were spindle-shaped and grew in a whorl pattern. Original magnification 100×, scale bars: 200 μm. (B) Flow cytometry detection of CD29, CD44, CD73 and CD105. The rate of positive expression for all factors was greater than 95%. (C) Detection of huMSC HLAII expression in PBMCs and huMSCs by western blot. No HLAII expression band was detected in the huMSCs. (D) Relative expression of HLAII in PBMCs and huMSCs. The bar graph shows no HLAII expression in huMSCs. (E) PCR amplification curves of HLA-DPA1 , HLA-DQA1 and HLA-DRA1 genes in PBMCs and huMSCs. (F) Relative expression of HLA-DPA1 , HLA-DQA1 and HLA-DRA1 genes in PBMCs and huMSCs. PCR results showed no expression of HLA-DPA1 , HLA-DQA1 or HLA-DRA1 genes in huMSCs. (G) Expression curves of serum IL-6, IL-10, IL-12, TGF-β, and TNF-α detected by ELISA method ( n = 5 rats per group). Compared with the TBI group, the serum pro-inflammatory cytokines IL-6, IL-12 and TNF-α in the Tail Vein and In Situ group were lower (### P < 0.001), whereas the inflammation inhibiting factors IL-10 and TGF-β were much higher (### P < 0.001), indicating that huMSCs exert good immunoregulatory effects. Data are expressed as the mean ± SD. *** P < 0.001, vs . PBMCs (one-way analysis of variance followed by least significant difference test). ELISA: Enzyme-linked immunosorbent assay; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; HLAII: human leukocyte antigen II; huMSCs: human umbilical cord mesenchymal stem cells; IL: interleukin; PBMCs: peripheral blood mononuclear cells; PCR: polymerase chain reaction; TBI: traumatic brain injury; TGF-β: transforming growth factor beta; TNF-α: tumor necrosis factor alpha.

Article Snippet: The sections were thereafter incubated overnight at 4°C with primary antibodies, including rabbit anti-human CD29 polyclonal antibody (Cat# PB9086, 1:100, Boster), proliferating cell nuclear antigen (PCNA) mouse monoclonal antibody (Cat# 2586S, 1:4000, CST, Danvers, MA, USA), Caspase 3 (CASP3) rabbit polyclonal antibody (Cat# 19677-1-AP, 1:200, Proteintech) and F4/80 rabbit polyclonal antibody (Cat# 28463-1-AP, 1:200, Proteintech), diluted in 1% bovine serum albumin containing PBS.

Techniques: Immunopeptidomics, Cell Culture, Flow Cytometry, Expressing, Western Blot, Amplification, Enzyme-linked Immunosorbent Assay, In Situ, Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Pre-clinical study of human umbilical cord mesenchymal stem cell transplantation for the treatment of traumatic brain injury: safety evaluation from immunogenic and oncogenic perspectives

doi: 10.4103/1673-5374.317985

Figure Lengend Snippet: Sites of accumulation of huMSCs in vivo in TBI rats . (A) Live imaging of the primary organs of experimental rats. (B, C) The average fluorescence intensity in the brain, liver and lung in the In Situ (B) and Tail Vein (C) groups. Data are expressed as the mean ± SD ( n = 5 rats at each time point). # P < 0.05, ## P < 0.01, vs . TBI group (one-way analysis of variance followed by least significant difference test). (D, E) CD29 (red, stained by CoraLite594) immunoreactivity in CFSE-labeled huMSCs in brain tissue (immunofluorescence staining, original magnification 200×, scale bars: 100 μm). CD29 and CFSE co-labeled huMSCs were identified in the In Situ group on days 1, 3 and 7 (D) in the brain lesions. A few CFSE-labeled huMSCs in the Tail Vein group were also observed in the brain lesions on days 1 and 3 (E). CFSE: Carboxyfluorescein succinimidyl ester; CON: TBI group; DAPI: 4′,6-diamidine-2′-phenylindole dihydrochloride; huMSCs: human umbilical cord mesenchymal stem cells; TBI: traumatic brain injury.

Article Snippet: The sections were thereafter incubated overnight at 4°C with primary antibodies, including rabbit anti-human CD29 polyclonal antibody (Cat# PB9086, 1:100, Boster), proliferating cell nuclear antigen (PCNA) mouse monoclonal antibody (Cat# 2586S, 1:4000, CST, Danvers, MA, USA), Caspase 3 (CASP3) rabbit polyclonal antibody (Cat# 19677-1-AP, 1:200, Proteintech) and F4/80 rabbit polyclonal antibody (Cat# 28463-1-AP, 1:200, Proteintech), diluted in 1% bovine serum albumin containing PBS.

Techniques: In Vivo, Imaging, Fluorescence, In Situ, Staining, Labeling, Immunofluorescence