Journal: Cell metabolism

Article Title: Reducing Hypothalamic Stem Cell Senescence Protects against Aging-Associated Physiological Decline.

doi: 10.1016/j.cmet.2020.01.002

Figure Lengend Snippet: Figure 4. Hnscr Binds to YB-1 and Protects It from Degradation (A) Mass spectrometry identified YB-1 in proteins pulled-down by biotinylated Hnscr in htNSCs. (B) Biotinylated Hnscr retrieved YB-1 in htNSCs as detected by western blot.

Article Snippet: To screen naturally occurring small-molecule compounds that target and stabilize YB-1, we performed molecular docking and virtual screening between YB-1 and the natural small molecular compounds library of Target Molecule (Target Mol), which contains more than 6,000 compounds.

Techniques: Mass Spectrometry, Western Blot

Journal: Cell metabolism

Article Title: Reducing Hypothalamic Stem Cell Senescence Protects against Aging-Associated Physiological Decline.

doi: 10.1016/j.cmet.2020.01.002

Figure Lengend Snippet: Figure 5. Overexpression of YB-1 Reduces the Senescent Effect of Hnscr Loss (A) The mRNA levels of Hnscr and p16INK4A in htNSCs. The value of the expression in the WT mice was set at an arbitrary value = 1. (B) The mRNA levels of YB-1 and p16INK4A in htNSCs. The value of the expression in the scramble-treated htNSCs was set at an arbitrary value = 1. (C) The mRNA levels of p16INK4A and Yb-1 in htNSCs. The value of the expression in the WT mice without YB-1 overexpression was set at an arbitrary value = 1. (D) p16INK4A and YB-1 protein levels in Hnscr-deleted or WT htNSCs with or without YB-1 overexpressed. (E) The levels of Hnscr and Yb-1 in htNSCs of WT mice 1 month after viral injection. The value of the expression in the control vector injection mice was set at an arbitrary value = 1. (F–H) Representative images (n = 18 photographs from 3 of experiments) of neurospheres generated from mice 1 month after viral injection (F). Scale bar, 100 mm. Their quantitation is shown in (G); relative size in (H). (legend continued on next page)

Article Snippet: To screen naturally occurring small-molecule compounds that target and stabilize YB-1, we performed molecular docking and virtual screening between YB-1 and the natural small molecular compounds library of Target Molecule (Target Mol), which contains more than 6,000 compounds.

Techniques: Over Expression, Expressing, Injection, Control, Plasmid Preparation, Generated, Quantitation Assay

Journal: Cell metabolism

Article Title: Reducing Hypothalamic Stem Cell Senescence Protects against Aging-Associated Physiological Decline.

doi: 10.1016/j.cmet.2020.01.002

Figure Lengend Snippet: Figure 6. Theaflavin 3-gallate (TF2A) Acts as an Hnscr Mimic (A) The optimized homology modeling structure of mouse YB-1. The green ribbon indicated a main barrel-like structure that is composed of 6 reverse-laminated beta-sheets. The upper part of the barrel-like structure (presented as the wheat-colored thread) indicated a dynamic loop area. Between them was the prediction of small molecule binding pockets (presented as pink balls) that consisted of 12 hydrophobic amino acids. (B) The molecular structure of TF2A. (C) The optimized binding modes with lowest binding energy generated by AutoDock Vina and key residues for interaction between TF2A and mouse YB-1. (D) The expression of YB-1 in mouse htNSCs treated with TF2A in a series of doses, as indicated. (E) The protein levels of YB-1 in TF2A pre-treated mouse htNSCs treated with cycloheximide (CHX). (F) Western blot analysis of YB-1-associated ubiquitination in TF2A pre-treated mouse htNSCs treated with Mg132. (G) Top: hPSCs derived neurospheres immunostained with Nestin and SOX2. Scale bar, 100 mm (n = 15 photographs from 3 experiments). Bottom: hPSC-derived NSCs were induced to differentiate after 7 days of differentiation. The cells were immunostained with neuronal marker Tuj1, oligodendrocyte marker O4, and astrocyte marker GFAP. Scale bar, 100 mm (n = 15 photographs from 3 experiments). (H) The protein levels of YB-1 in hPSC-derived NSCs treated with TF2A in a series of dose as indicated. Top: representative images of western blot. Bottom: quantification analysis of YB-1 in relation to GAPDH. Data are shown as mean ± SEM. The cell experiments were repeated 3 times. *p < 0.05; **p < 0.01; ***p < 0.001 by two-tailed Student’s t test or one-way ANOVA.

Article Snippet: To screen naturally occurring small-molecule compounds that target and stabilize YB-1, we performed molecular docking and virtual screening between YB-1 and the natural small molecular compounds library of Target Molecule (Target Mol), which contains more than 6,000 compounds.

Techniques: Binding Assay, Generated, Expressing, Western Blot, Ubiquitin Proteomics, Derivative Assay, Marker, Two Tailed Test

Journal: The Journal of infectious diseases

Article Title: Aberrant Humoral Immune Responses in Neurosyphilis: CXCL13/CXCR5 Play a Pivotal Role for B-Cell Recruitment to the Cerebrospinal Fluid.

doi: 10.1093/infdis/jix233

Figure Lengend Snippet: Figure 1. Enrichment of CD19+ B cells and memory B cells in the CSF of NS patients. A, Gating strategies. B, Representative flow cytometry analysis of the frequency of CSF total CD19+ B cells and subsets of CD19+ B cells between non-NS and NS patients (first row). Bar charts of the frequency of B-cell subsets of patients with asymptomatic and symptomatic NS (second row). Comparison of the proportion of CSF total CXCR5+ B cells in different groups (third row). C, Serial changes of CSF total CD19+ B cells in the patient with NS following first and second treatment. D, Correlation of frequency of CD19+ B cells and CSF protein levels, CSF WBC counts, or CSF VDRL titer. Abbreviations: CSF, cerebrospinal fluid; NS, neurosyphilis; VDRL, Venereal Disease Research Laboratory; WBC, white blood cell.

Article Snippet: Paraffin-embedded brain biopsy (4 μm) was stained with appropriate concentration of primary antibodies against CXCL13, CXCR5, CD20, CD3, CD35, CD138 (Maixin-Bio, Fujian, China), and spirochete (Biocare Medical, Pacheco, CA), then incubated with streptavidin-biotin complex system according to manufacturer’s instructions (Boster, Pleasanton, CA).

Techniques: Flow Cytometry, Comparison

Journal: The Journal of infectious diseases

Article Title: Aberrant Humoral Immune Responses in Neurosyphilis: CXCL13/CXCR5 Play a Pivotal Role for B-Cell Recruitment to the Cerebrospinal Fluid.

doi: 10.1093/infdis/jix233

Figure Lengend Snippet: Figure 5. Pathologic diagnosis of intracranial syphilitic gumma and cellular organization of follicle-like structures. HE staining of intracranial syphilitic gumma (first row). Immunostaining of spirochete in intracranial syphilitic gumma (second row). Immunostaining of CXCL13 in intracranial syphilitic gumma and follicle-like structure (third row). Immunostaining of CXCR5 in intracranial syphilitic gumma and follicle-like structure (fourth row). Immunostaining of CD20+ B cells in intra- cranial syphilitic gumma and follicle-like structure (fifth row). Immunostaining of CD3+ T cells in intracranial syphilitic gumma and follicle-like structure (sixth row). Immunostaining of CD35+ FDCs in intracranial syphilitic gumma and follicle–like structure (seventh row). Immunostaining of CD138+ plasma cells in intracranial syphilitic gumma and follicle-like structure (eighth row). Abbreviations: FDCs, follic- ular dendritic cells; HE, hematoxylin and eosin.

Article Snippet: Paraffin-embedded brain biopsy (4 μm) was stained with appropriate concentration of primary antibodies against CXCL13, CXCR5, CD20, CD3, CD35, CD138 (Maixin-Bio, Fujian, China), and spirochete (Biocare Medical, Pacheco, CA), then incubated with streptavidin-biotin complex system according to manufacturer’s instructions (Boster, Pleasanton, CA).

Techniques: Biomarker Discovery, Staining, Immunostaining, Clinical Proteomics

Journal: Frontiers in Pharmacology

Article Title: Mechanism of Paeoniflorin on ANIT-Induced Cholestatic Liver Injury Using Integrated Metabolomics and Network Pharmacology

doi: 10.3389/fphar.2021.737630

Figure Lengend Snippet: Effect of PF on the expression of CYP2C9, ABCB1, MAOB, CDC25B, and MTOR in the treatment of cholestatic liver injury detected by western blotting. (A) Western blotting images of CYP2C9, ABCB1, MAOB, CDC25B, and MTOR; (B) Relative protein expression of CDC25B; (C) Relative protein expression of CYP2C9; (D) Relative protein expression of MAOB; (E) Relative protein expression of ABCB1; (F) Relative protein expression of MTOR. The data are expressed as the mean ± SD, n = 3. ## p < 0.01, # p < 0.05 compared with the control group, ** p < 0.01, * p < 0.05 compared with ANIT group.

Article Snippet: The PVDF membranes were blocked with 5% fat-free milk at room temperature for 2 hours, then incubated overnight at 4°C with antibodies against anit-CDC25B (PB9488, BOSTER, dilution: 1:1,000), anit-MTOR (A00003-2, BOSTER, dilution: 1:1,000), anit-CYP2C9 (16546-1-AP, proteintech, dilution: 1:2,000), anit-ABCB1 (A00049-3, BOSTER, dilution: 1:1,000), anit-MAOB (PB9665, BOSTER, dilution: 1:1,000).

Techniques: Expressing, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Mechanism of Paeoniflorin on ANIT-Induced Cholestatic Liver Injury Using Integrated Metabolomics and Network Pharmacology

doi: 10.3389/fphar.2021.737630

Figure Lengend Snippet: Effect of PF on the expression of CYP2C9, ABCB1, MAOB, CDC25B, and MTOR in the treatment of cholestatic liver injury. (A) The expression of CYP2C9 in the liver tissue; (B) The expression of ABCB1 in the liver tissue; (C) The expression of MAOB in the liver tissue; (D) The expression of CDC25B in the liver tissue; (E) The expression of MTOR in the liver tissue. The data are expressed as the mean ± SD, n = 3. ## p < 0.01, # p < 0.05 compared with the control group, ** p < 0.01, * p < 0.05 compared with ANIT group.

Article Snippet: The PVDF membranes were blocked with 5% fat-free milk at room temperature for 2 hours, then incubated overnight at 4°C with antibodies against anit-CDC25B (PB9488, BOSTER, dilution: 1:1,000), anit-MTOR (A00003-2, BOSTER, dilution: 1:1,000), anit-CYP2C9 (16546-1-AP, proteintech, dilution: 1:2,000), anit-ABCB1 (A00049-3, BOSTER, dilution: 1:1,000), anit-MAOB (PB9665, BOSTER, dilution: 1:1,000).

Techniques: Expressing, Control