Journal: Cell reports

Article Title: Selenoprotein P is a target for regulating extracellular vesicle biogenesis and secretion from activated microglia in vivo

doi: 10.1016/j.celrep.2024.115025

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: pLV-EF1α-mEmerald-CD9-miR9T , Tsuneya Ikezu ( ) , Addgene: Cat#170452.

Techniques: Virus, shRNA, Control, Recombinant, Staining, RNA Sequencing, Software, Saline, Imaging

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: TREM2 promotes cholesterol uptake and foam cell formation in atherosclerosis

doi: 10.1007/s00018-023-04786-9

Figure Lengend Snippet: TREM2 expression is increased in atherosclerotic plaques. a The mRNA levels of TREM2 in human atherosclerotic plaque and non-carotid plaque vascular tissue (adjacent carotid tissue). The original data came from the GEO database GSE43292. student’s t test. b TREM2 mRNA was detected in the aorta from ApoE−/− mice fed a HFD for different periods (week; n = 5/group, one-way ANOVA test followed by a post hoc Tukey’s test). c Graph shows the trend of TREM2 mRNA expression after stimulation with oxLDL (50 µg/mL) for different times (n = 4/group, one-way ANOVA followed by a post hoc Tukey’s test). d, e Representative confocal images of TREM2 (red) in the atherosclerotic plaques of the aortic sinus from ApoE−/− mice in the indicated groups (n = 6/group). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue). Scale bar: 50 µm (upper panel), 25 µm (lower panel). One-way ANOVA test followed by a post hoc Tukey’s test. All data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, ****P < 0.0001; ns, P > 0.05. AS atherosclerosis, H&E staining hematoxylin–eosin staining

Article Snippet: TREM2-knockout mice ( Trem2 −/− ) on the C57BL/6N background were purchased from Cyagen (Santa Clara, CA, USA, https://www.cyagen.com/cn/zh-cn/sperm-bank-live/83433 ).

Techniques: Expressing, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: TREM2 promotes cholesterol uptake and foam cell formation in atherosclerosis

doi: 10.1007/s00018-023-04786-9

Figure Lengend Snippet: TREM2 is expressed in foam cells derived from macrophages and SMCs in plaques. a–c Representative images of aortic sinuses obtained from the Trem2+/+/ApoE−/− and Trem2−/−/ApoE−/− groups co-stained TREM2 (red) with BODIPY (a, green), CD68 (b, green), and α-SMA (c, green). For fluorescence images, nuclei were stained with DAPI (blue). d–f The number of TREM2/BODIPY- (d), TREM2/CD68-, (e) or TREM2/α-SMA-positive cells (f) per mm2 of plaques in the indicated groups is shown (n = 6–7/group, one-way ANOVA followed by a post hoc Tukey’s test). All data are expressed as the mean ± SEM from three to five independent experiments. Scale bar: 25 μm. ***P < 0.001, ****P < 0.0001

Article Snippet: TREM2-knockout mice ( Trem2 −/− ) on the C57BL/6N background were purchased from Cyagen (Santa Clara, CA, USA, https://www.cyagen.com/cn/zh-cn/sperm-bank-live/83433 ).

Techniques: Derivative Assay, Staining, Fluorescence

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: TREM2 promotes cholesterol uptake and foam cell formation in atherosclerosis

doi: 10.1007/s00018-023-04786-9

Figure Lengend Snippet: Deletion of TREM2 ameliorates plaque formation and lipid deposition. a Trem2+/+/ApoE−/− and Trem2−/−/ApoE−/− mice were fed a HFD for 12 weeks. The percentage of whole aorta and aortic arch area that was occupied by the ORO-positive area was assessed (n = 6–7/group, student’s t test). Scale bar: 0.5 cm. b Images of H&E, ORO, and BODIPY-stained tissues showing the aortic sinus level obtained from mice. The red area shows plaque lipid in the aorta. Representative images of the aortic sinus were obtained from mice in the Trem2+/+/ApoE−/− and Trem2−/−/ApoE−/− groups. For H&E staining, the necrotic core was marked by a yellow line and zoomed in locally. Scale bar: 100 μm. c Representative images showing plaque area, the percentage of the plaque area of the necrotic core and ORO-positive area (n = 6/groups, student’s t test). The foam cell number of the plaque area occupied by BODIPY is presented among the indicated groups (green; n = 6/group, student’s t test). DAPI was used to stain nuclei (blue). All data are expressed as the mean ± SEM from three to five independent experiments. *P < 0.05, **P < 0.01, ****P < 0.0001

Article Snippet: TREM2-knockout mice ( Trem2 −/− ) on the C57BL/6N background were purchased from Cyagen (Santa Clara, CA, USA, https://www.cyagen.com/cn/zh-cn/sperm-bank-live/83433 ).

Techniques: Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: TREM2 promotes cholesterol uptake and foam cell formation in atherosclerosis

doi: 10.1007/s00018-023-04786-9

Figure Lengend Snippet: TREM2 deficiency decreases foam cell formation in vivo. a Immunoblot analysis of the expression of p-p38, t-p38, p-PPARγ, t-PPARγ, and CD36 in extracts from the aortas of mice in the indicated group. Trem2+/+/ApoE−/− mice and Trem2−/−/ApoE−/− mice were fed a HFD for 12 weeks to induce atherosclerotic lesions. Quantitative data represent the fold change observed. Protein expression levels are calculated in (b n = 6/group, student’s t test). c Fluorescence images of CD68-BODIPY or α-SMA-BODIPY at the aortic sinus from Trem2+/+/ApoE−/− mice and Trem2−/−/ApoE−/− mice. Nuclei were stained with DAPI (blue). Scale bar: 50 μm. d, e The number of CD68/BODIPY- or α-SMA/BODIPY-positive cells per mm2 of plaques in the indicated groups is presented (n = 6/group, student’s t test). All data are shown as the mean ± SEM from three to five independent experiments. *P < 0.05, **P < 0.01

Article Snippet: TREM2-knockout mice ( Trem2 −/− ) on the C57BL/6N background were purchased from Cyagen (Santa Clara, CA, USA, https://www.cyagen.com/cn/zh-cn/sperm-bank-live/83433 ).

Techniques: In Vivo, Western Blot, Expressing, Fluorescence, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: TREM2 promotes cholesterol uptake and foam cell formation in atherosclerosis

doi: 10.1007/s00018-023-04786-9

Figure Lengend Snippet: TREM2 promotes foam cell formation in vitro. Lentiviral vectors were constructed to deliver the TREM2 gene (LV-TREM2). Lentiviral GFP (LV-GFP) was used as the control group (LV-GFP). a, b Images of ORO-stained ox-LDL-loaded Raw264.7 cells and mSMCs (a) in the indicated group. Graphs (b) show the ORO area positive per cell (n = 9/group, student’s t test). Scale bar: 10 μm. c, d Representative Western blots (left) of extracts obtained from Raw264.7 cells and mSMCs among the indicated groups of LV-GFP versus LV-TREM2 (n = 6/group, student’s t test). e, f Quantitative real-time PCR analysis of the mRNA levels of CD36 in macrophages and mSMCs in the LV-GFP or LV-TREM2 group. Data are normalized to ACTB expression (n = 5–6/group, student’s t test). g, Western blotting and statistics of BMDMs and mSMCs isolated from Trem2+/+ and Trem2−/− mice after stimulation with oxLDL at 50 µg/mL (n = 6/group, student’s t test). All data are expressed as the mean ± SEM from three to five independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Article Snippet: TREM2-knockout mice ( Trem2 −/− ) on the C57BL/6N background were purchased from Cyagen (Santa Clara, CA, USA, https://www.cyagen.com/cn/zh-cn/sperm-bank-live/83433 ).

Techniques: In Vitro, Construct, Staining, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Isolation

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: TREM2 promotes cholesterol uptake and foam cell formation in atherosclerosis

doi: 10.1007/s00018-023-04786-9

Figure Lengend Snippet: TREM2 stimulates cholesterol uptake by regulating CD36. a Representative fluorescence images of Raw264.7 cells and mSMCs labeled with DiI-oxLDL at 37 °C for 4 h. Scale bar: 10 μm. b The mean DiI-oxLDL burden detected by flow cytometry (n = 9/group, student’s t-test). c and e Optical microscopy images (top) or confocal laser scanning microscopy images (bottom) of Raw264.7 cells and mSMCs labeled by ORO or DiI-oxLDL. CD36 small-interfering RNA (siRNA)-transfected Raw264.7 cells and mSMCs in the LV-GFP and LV-TREM2 groups before stimulation with oxLDL or DiI-oxLDL. Scale bars: 10 μm. Graphs (d, f) show the area positive for ORO per cell or mean DiI-oxLDL burden (mFI, n = 6 and 9/group, respectively; two-way ANOVA test followed by a post hoc Tukey’s test). All data are expressed as the mean ± SEM from 3 to 5 independent experiments. *P < 0.05, ***P < 0.001, ****P < 0.0001. mFI mean fluorescence intensity

Article Snippet: TREM2-knockout mice ( Trem2 −/− ) on the C57BL/6N background were purchased from Cyagen (Santa Clara, CA, USA, https://www.cyagen.com/cn/zh-cn/sperm-bank-live/83433 ).

Techniques: Fluorescence, Labeling, Flow Cytometry, Microscopy, Confocal Laser Scanning Microscopy, Small Interfering RNA, Transfection

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: TREM2 promotes cholesterol uptake and foam cell formation in atherosclerosis

doi: 10.1007/s00018-023-04786-9

Figure Lengend Snippet: TREM2 regulates MAPK-p38 and PPARγ phosphorylation. a–d Representative Western blots and normalized ratios of p-p38:t-p38, p-JNK1/2:t-JNK1/2, p-ERK1/2:t-ERK1/2, and p-PPARγ:t-PPARγ in RAW264.7 cells or mSMCs infected with LV-GFP or LV-TREM2 group (n = 6/group, student’s t test). e, f Representative images of ORO- and DiI-oxLDL-positive areas in Raw264.7 cells or mSMCs obtained from the indicated group. Nuclei were stained with DAPI (blue). RAW264.7 cells and mSMCs were cultured with the p38 phosphorylation agonist dehydrocorydaline (Deh) before oxLDL or DiI-oxLDL stimulation, and then the ORO- or DiI-oxLDL-positive area was analyzed (n = 6–9/group, two-way ANOVA followed by a post hoc Tukey’s test). g, h Immunoblot analysis of the expression of p-p38, t-p38, p-PPARγ, t-PPARγ, and CD36 in RAW264.7 cells and mSMCs treated with dehydrocorydaline (Deh) or an equal volume of DMSO (ctr, n = 6/group, two-way ANOVA test followed by a post hoc Tukey’s test). Quantitative data represent the fold change observed after normalizing the indicated protein band intensities to GAPDH. All data are presented as the mean ± SEM from three to five independent experiments. Scale bar: 10 μm. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. mFI mean fluorescence intensity. Deh, dehydrocorydaline

Article Snippet: TREM2-knockout mice ( Trem2 −/− ) on the C57BL/6N background were purchased from Cyagen (Santa Clara, CA, USA, https://www.cyagen.com/cn/zh-cn/sperm-bank-live/83433 ).

Techniques: Western Blot, Infection, Staining, Cell Culture, Expressing, Fluorescence

Journal: Microbiology Spectrum

Article Title: Amuvatinib Blocks SARS-CoV-2 Infection at the Entry Step of the Viral Life Cycle

doi: 10.1128/spectrum.05105-22

Figure Lengend Snippet: Amuvatinib displays broad-spectrum antiviral activity against SARS-CoV-2 variants. (A) HEK293T cells were cotransfected with ACE2 and TMPRSS2 for 20 h and then treated with either DMSO or 20 μM amuvatinib for 10 min. Cells were infected with wild-type SARS-CoV-2pp in the absence or presence of 20 μM amuvatinib for 1 h, and then culture media were replaced with fresh media without amuvatinib. At 36 h postinfection, the cells were harvested, and viral infection was determined by luciferase assay. (B to F) HEK293T cells were treated as described in panel A using Alpha (B), Beta (C), Gamma (D), Delta (E), and Omicron (F) variants of SARS-CoV-2pp. All experiments were performed in triplicate, and data represent averages. **, P < 0.01; ***, P < 0.001; Wild-type, Wuhan-Hu-1 (2019-nCoV).

Article Snippet: Cells were transfected with 1 μg of SARS-CoV-2 spike expression plasmid, 3.1 μg of Gag-Pol packaging plasmid, and 3.1 μg of transfer vector encoding the firefly luciferase reporter protein using polyethyleneimine to create wild-type ( https://www.addgene.org/149539/ ), Alpha ( https://www.addgene.org/170451 ), Beta ( https://www.addgene.org/170449 ), Gamma ( https://www.addgene.org/170450 ), Delta ( https://www.addgene.org/172320 ), and Omicron ( https://www.addgene.org/180375 ) variants of SARS-CoV-2pp.

Techniques: Activity Assay, Infection, Luciferase