Journal: Cell Stem Cell

Article Title: Selective advantage of mutant stem cells in human clonal hematopoiesis is associated with attenuated response to inflammation and aging

doi: 10.1016/j.stem.2024.05.010

Figure Lengend Snippet:

Article Snippet: APC anti-human CD271 (clone ME20.4-1.H4) , Miltenyi , Cat# 130-113-418; RRID: AB_2733363.

Techniques: Recombinant, Reverse Transcription, DNA Library Preparation, DNA Sequencing, RNA Sequencing, Purification, Sequencing, Software, Sterility

Journal: Journal of Clinical Medicine

Article Title: Moderate Aerobic Exercise Regulates Follicular Dysfunction by Initiating Brain-Derived Neurotrophic Factor (BDNF)-Mediated Anti-Apoptotic Signaling Pathways in Polycystic Ovary Syndrome

doi: 10.3390/jcm11195584

Figure Lengend Snippet: Sequences of primers designed for RT-qPCR.

Article Snippet: Sections were incubated overnight at 4 °C with antibodies against BDNF (1:100, Wanleibio), TrkB (1:100, Wanleibio), p75NTR (1:100, 55014-1-AP, Proteintech, Chicago, IL, USA), and Cleaved-Caspase-3 (1:100, 66470-2-Ig, Proteintech) at a 1:100 dilution.

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Moderate Aerobic Exercise Regulates Follicular Dysfunction by Initiating Brain-Derived Neurotrophic Factor (BDNF)-Mediated Anti-Apoptotic Signaling Pathways in Polycystic Ovary Syndrome

doi: 10.3390/jcm11195584

Figure Lengend Snippet: The expression of BDNF, TrkB, and p75NTR in ovarian follicles of PCOS rats after aerobic exercise. Rats received DHEA for the induction of polycystic ovarian syndrome, together with or without exercise treatment. ( A , B ) The expression levels of BDNF in ovarian follicles at different stages were analyzed by immunohistochemistry. ( C , D ) The expression levels of BDNF in ovarian follicles were analyzed by immunofluorescence. ( E , F ) The co-expression of TrkB and p75NTR in ovarian follicles at different stages was detected by immunofluorescence double labeling. n = 10 in each group. Data are shown as mean ± SEM. ** p < 0.05, vs. control group; ## p < 0.01, vs. PCOS group.

Article Snippet: Sections were incubated overnight at 4 °C with antibodies against BDNF (1:100, Wanleibio), TrkB (1:100, Wanleibio), p75NTR (1:100, 55014-1-AP, Proteintech, Chicago, IL, USA), and Cleaved-Caspase-3 (1:100, 66470-2-Ig, Proteintech) at a 1:100 dilution.

Techniques: Expressing, Immunohistochemistry, Immunofluorescence, Labeling, Control

Journal: Journal of Clinical Medicine

Article Title: Moderate Aerobic Exercise Regulates Follicular Dysfunction by Initiating Brain-Derived Neurotrophic Factor (BDNF)-Mediated Anti-Apoptotic Signaling Pathways in Polycystic Ovary Syndrome

doi: 10.3390/jcm11195584

Figure Lengend Snippet: Aerobic exercise activated BDNF signaling in the ovary of DHEA-induced PCOS rats. Rats received DHEA for the induction of polycystic ovarian syndrome, together with or without exercise treatment. ( A ) mRNA expression of BDNF, TrkB, and p75NTR factors in ovarian tissue was analyzed by real-time PCR. ( B , C ) The expression of BDNF, TrkB, and p75NTR in ovarian tissue was assessed by Western blot assay. ( D , E ) The expression of PI3K, AKT, and p-AKT in ovarian tissue was assessed by Western blot assay. ( F – I ) The expression of p75NTR, NF-κB, p-NF-κB, JNK, and p-JNK in ovarian tissue was assessed by Western blot assay. n = 10 in each group. Data are shown as mean ± SEM. ** p < 0.05, vs. control group; ## p < 0.01, vs. PCOS group.

Article Snippet: Sections were incubated overnight at 4 °C with antibodies against BDNF (1:100, Wanleibio), TrkB (1:100, Wanleibio), p75NTR (1:100, 55014-1-AP, Proteintech, Chicago, IL, USA), and Cleaved-Caspase-3 (1:100, 66470-2-Ig, Proteintech) at a 1:100 dilution.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Steady-state distribution and delivery to the plasma membrane of PLAP and NTR-PLAP in transfected FRT cells. (A) Schemes of PLAP and NTR-PLAP (a fusion protein containing the ectodomain of p75NTR and the C-terminal GPI attachment signal of PLAP). (B) FRT cells stably expressing PLAP or NTR-PLAP were grown to confluence and pulsed overnight with [35S]met–cys or [35S]cys, respectively. After surface biotinylation from the apical (Ap) and basolateral (BL) domains, cells were lysed, immunoprecipitated against specific antibodies, and reprecipitated against streptavidin. (C) FRT cells were labeled for 20 min with [35S]met–cys or [35S]cys. At different times of the chase, surface proteins were biotinylated from the apical (Ap) or the basolateral (BL) side. Cells were then lysed and immunoprecipitated with antibodies against PLAP or NTR and with streptavidin beads. Samples were run on 10% SDS-PAGE and detected by fluorography. PLAP and NTR-PLAP are directly sorted to the apical surface in FRT cells, whereas a small percentage of the two proteins is delivered with similar kinetics to the basolateral domain.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Transfection, Stable Transfection, Expressing, Immunoprecipitation, Labeling, SDS Page

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Pulse-chase analysis and purification on sucrose gradients of transmembrane forms of PLAP and p75NTR in transfected FRT cells. (A) Schemes of p75NTR and PLAP-PS321 (a fusion protein containing the ectodomain of PLAP and the transmembrane and the cytoplasmic domain of the PS321 NTR mutant). (B) Pulse-chase and TX-100 extraction of p75NTR and PLAP-PS321. FRT cells expressing p75NTR and PLAP-PS321 were labeled with [35S]cys or [35S]met–cys, respectively, and subsequently treated as described in Figure ​Figure2A.2A. (C) Purification of p75NTR- and PLAP-PS321–enriched fractions on sucrose density gradients. Labeled proteins of FRT cells expressing p75NTR and PLAP-PS321 were solubilized in TX-100 and purified as described in Figure ​Figure2B.2B. Both transmembrane proteins are soluble in TX-100 and do not float on sucrose density gradients.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Pulse Chase, Purification, Transfection, Mutagenesis, Expressing, Labeling

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Surface expression of GPI-anchored, secretory, and transmembrane proteins after FB1 treatment. (A) FRT cells stably expressing PLAP, NTR-PLAP, or gD1-DAF were grown to confluence in the absence (−) or presence (+) of FB1, as described in MATERIALS AND METHODS. After 30 min of labeling with [35S]met–cys or [35S]cys and a 2-h chase, cells were surface biotinylated from the apical (Ap) or the basolateral (BL) domain, lysed, immunoprecipitated against specific antibodies, and reprecipitated against streptavidin. (B) Amounts of labeled proteins were quantified from three independent experiments and are shown as means ± SD. Hatched bars represent apical proteins, and black bars represent basolateral proteins. Apical PLAP and NTR-PLAP, but not basolateral gD1-DAF, became unpolarized after FB1 treatment. (C) FRT cells expressing PLAP-sec, NTR-sec, p75NTR, or PLAP-PS321 were grown to confluence in the absence (−) or presence (+) of FB1, as described in MATERIALS AND METHODS. After labeling with [35S]met–cys or [35S]cys for 20 min and a chase of 2 h, apical and basolateral media were collected separately and cells were surface biotinylated from apical or basolateral domains. Polarized secretion of the secretory forms was revealed by immunoprecipitation in the apical and basolateral medium (top two panels), whereas biotinylated transmembrane proteins were revealed by double immunoprecipitation with specific antibody and streptavidin beads (bottom two panels). (D) Amounts of labeled proteins were quantified from three independent experiments and are shown as means ± SD. Hatched bars represent apical proteins, and black bars represent basolateral proteins. FB1 does not alter the sorting of transmembrane and secretory proteins.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Expressing, Stable Transfection, Labeling, Immunoprecipitation

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Surface expression of GPI-anchored and transmembrane proteins after cholesterol depletion. (A) FRT cells expressing PLAP, NTR-PLAP, gD1-DAF, p75NTR, or PLAP-PS321 were grown on filters and treated with mevinolin (Mev.; 10 μM) for 48 h and M-β-CD (10 mM) for 60 min. Cells were then pulse labeled for 30 min, chased for 2 h, and biotinylated as usual. (B) Amounts of labeled proteins were quantified in three independent experiments and are shown as means ± SD. Hatched bars represent apical (Ap) proteins, and black bars represent basolateral (BL) proteins. Removal of cholesterol does not alter the surface expression of apical and basolateral GPI-anchored and transmembrane proteins.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Expressing, Labeling

Journal: Purinergic Signalling

Article Title: Up-regulation of P2X7 receptors mediating proliferation of Schwann cells after sciatic nerve injury

doi: 10.1007/s11302-015-9445-8

Figure Lengend Snippet: Expression of P2X7R ir in longitudinal sections of normal sciatic nerves. a–c show co-localization of P2X7R ir (red) and S100Aβ ir (green). Note an arrowhead showing a fibre-like structure and an arrow showing a trapezoid structure in a and b; c is the merged image of a and b. Note an arrow indicating a trapezoid structure double labelled by both P2X7R and S100β antibodies (yellow). d–f show co-localization of P2X7R ir (red) and Tuj-1 ir (green). Note an arrow showing a trapezoid structure in d and an arrow showing an axon in e. f is the merged image of d and f; note an arrow indicating a green axon passing through the middle of five trapezoid structures. g–i show co-localization of P2X7R ir (red) and p75NTR ir (green). Note an arrow showing a fibre-like structure in g and h. i is the merged image of g and h. An arrow indicates a double-labelled non-myelinating Schwann cell with P2X7R and p75NTR antibodies (yellow). j–l show co-localization of P2X7R ir (red) and MBP ir (green). Note an arrow showing a fibre-like structure with P2X7R ir, which was not labelled by MBP in l. m–o show co-localization of P2X7R ir (red) and CASPR ir (green). Note that no colocalization of P2X7R ir and CASPR ir was observed. Scale bars in a–c and g–i = 100 μm; scale bars in d–f = 50 μm

Article Snippet: The sections were washed 3–5 min in PBS, and then preincubated in a blocking solution (10 % normal bovine serum, 0.2 % Triton X-100, 0.4 % sodium azide in 0.01 mol/L PBS, pH 7.2) for 30 min followed by incubation with the primary antibodies: P2X7R (1:1000), Alomone, rabbit polyclonal, APR-004; S100β (1:400), Tuj-1 (1:200), p75NTR, PCNA (1:200), MBP (1:200), mouse monoclonal antibodies from Boster, CASPR (1:400) mouse monoclonal antibodies from Santa Cruz, at room temperature overnight.

Techniques: Expressing