antibody against cxcl13  (R&D Systems)

Journal: Cell and tissue research

Article Title: CXCL13 is expressed in a subpopulation of neuroendocrine cells in the murine trachea and lung.

doi: 10.1007/s00441-021-03552-2

Figure Lengend Snippet: Fig. 4 CXCL13 is expressed in two-thirds of the murine tracheal neuroendocrine cells. a, b Immunohistochemistry of tracheal whole mounts and the corresponding quantification of their immunoreactive cells; maximum intensity projec- tions of z-stacks of confocal optical sections. a Immuno- histochemistry with antibodies against CXCL13 (a) (green) and PGP9.5 (a′) (red), labeling single neuroendocrine cells and nerve fibers. CXCL13+/ PGP9.5+ cells are indicated by arrowheads; CXCL13−/ PGP9.5+ cells are indicated by ( <). Data points in the scatter plot (a‴) represent mean values of counts in one trachea (n = 5 tracheas); mean and SEM are indicated. The pie chart shows the percentages of CXCL13+/ PGP9.5+ and CXCL13−/ PGP9.5+ cells (n = 2254 cells pooled from 5 tracheas). b Immunohistochemistry with antibodies against CXCL13 (b) (green) and CGRP (b′) (red), labeling single neu- roendocrine cells and nerve fibers. CXCL13+/CGRP+ cells are indicated by arrowheads; CXCL13−/CGRP+ cells are indicated by ( <); CXCL13+/ CGRP− cells are indicated by (*). Data points in the scatter plot (b‴) represent mean values of counts in one trachea (n = 5 tracheas); mean and SEM are indicated. The pie chart shows the percentages of phenotypes (n = 2650 cells pooled from 5 tracheas)

Article Snippet: Floating sections were rinsed in PBS, and unspecific protein binding sites were saturated with 10% normal porcine serum in 0.005 M PBS for 1 h. Sections were incubated overnight with goat polyclonal antibody against CXCL13 (1:400 AF470, R&D Systems) or rabbit polyclonal antibody against αCGRP (1:20,000; T-4032, Peninsula Laboratories) followed by incubation for 1 h with peroxidase-conjugated pig anti-rabbit Ig (1:100; P0217, Dako, Santa Clara, USA) to detect CGRP or with biotinylated secondary donkey anti-goat IgG (1:400; 705–065- 147, Dianova) to detect CXCL13.

Techniques: Immunohistochemistry, Labeling

Journal: Cell and tissue research

Article Title: CXCL13 is expressed in a subpopulation of neuroendocrine cells in the murine trachea and lung.

doi: 10.1007/s00441-021-03552-2

Figure Lengend Snippet: Fig. 6 CXCL13 is less expressed in murine broncho- pulmonary solitary and clustered neuroendocrine cells. Immunohistochemistry of lung cryosections with antibodies against CXCL13 (orange) and CGRP (green) and the relative frequencies of immunoreactive phenotypes. a Solitary neuroen- docrine cell co-labeled with antibodies against CXCL13 and CGRP. b Solitary neu- roendocrine cell only labeled with antibodies against CGRP. c A cluster of neuroendocrine cells (neuroepithelial body) consisting of more than 7 cells, 2 of them are co-labeled with antibodies against CXCL13 and CGRP. d Pie chart shows percentages of CXCL13+/ CGRP+ and CXCL13−/CGRP+- immunolabeled cells in solitary neuroendocrine cells (n = 73 cells pooled from 5 animals). e Pie chart shows the percent- age of CXCL13+/CGRP+ and CXCL13−/CGRP+- immunolabeled cells in neu- roepithelial bodies (n = 1475 cells pooled from 5 animals)

Article Snippet: Floating sections were rinsed in PBS, and unspecific protein binding sites were saturated with 10% normal porcine serum in 0.005 M PBS for 1 h. Sections were incubated overnight with goat polyclonal antibody against CXCL13 (1:400 AF470, R&D Systems) or rabbit polyclonal antibody against αCGRP (1:20,000; T-4032, Peninsula Laboratories) followed by incubation for 1 h with peroxidase-conjugated pig anti-rabbit Ig (1:100; P0217, Dako, Santa Clara, USA) to detect CGRP or with biotinylated secondary donkey anti-goat IgG (1:400; 705–065- 147, Dianova) to detect CXCL13.

Techniques: Immunohistochemistry, Labeling, Immunolabeling

Journal: Journal of Neuroinflammation

Article Title: Chemokine CXCL13 acts via CXCR5-ERK signaling in hippocampus to induce perioperative neurocognitive disorders in surgically treated mice

doi: 10.1186/s12974-020-02013-x

Figure Lengend Snippet: Schematic illustration of the experimental design. CXCL13 C-X-C motif chemokine 13, CXCR5 C-X-C motif chemokine receptor 5, HE hematoxylin-eosin staining, i.c.v. intracerebroventricular, IL - 1β interleukin-1 beta, TNF - α tumor necrosis factor alpha, WT wide-type

Article Snippet: Membranes were blocked, then incubated overnight at 4 °C with antibodies against CXCL13 (1:100; Novus Biologicals, Littleton, CO, USA) or CXCR5 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA); or antibodies against ERK (1:500), p-ERK (1:500), IL-1β (1:1000), or TNF-α (1:500) (Cell Signaling, Beverly, MA, USA).

Techniques: Staining

Journal: Journal of Neuroinflammation

Article Title: Chemokine CXCL13 acts via CXCR5-ERK signaling in hippocampus to induce perioperative neurocognitive disorders in surgically treated mice

doi: 10.1186/s12974-020-02013-x

Figure Lengend Snippet: Surgery up-regulates CXCL13 in mouse hippocampus. a Relative level of CXCL13 mRNA was measured by RT-qPCR in wild type mice at 1, 3, and 7 days after surgery. b Western blotting analysis of CXCL13 at 7 days after surgery. β-actin was used as an internal control ( n = 8 per group). c Representative images of CXCL13 immunohistochemistry in the hippocampal CA1, CA3, and dentate gyrus regions. Positively stained cells are brown (red arrow). Magnification, × 5 for total hippocampus; × 20 for regions CA1, CA3, and DG in hippocampus. All experiments were performed with n = 4 per group. * P < 0.05 vs. control. CXCL13 C-X-C motif chemokine 13, DG dentate gyrus, RT - qPCR reverse transcription-quantitative polymerase chain reaction

Article Snippet: Membranes were blocked, then incubated overnight at 4 °C with antibodies against CXCL13 (1:100; Novus Biologicals, Littleton, CO, USA) or CXCR5 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA); or antibodies against ERK (1:500), p-ERK (1:500), IL-1β (1:1000), or TNF-α (1:500) (Cell Signaling, Beverly, MA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Control, Immunohistochemistry, Staining, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Journal of Neuroinflammation

Article Title: Chemokine CXCL13 acts via CXCR5-ERK signaling in hippocampus to induce perioperative neurocognitive disorders in surgically treated mice

doi: 10.1186/s12974-020-02013-x

Figure Lengend Snippet: CXCL13 knockdown alleviates PND-like cognitive deficits in mice. Mice were pretreated with CXCL13 shRNA or scrambled shRNA, subjected to surgery and then behavioral tests were performed 7 days after surgery. a Western blotting and densitometry of hippocampal CXCL13, as well as b RT-qPCR analysis of the corresponding mRNA. c Time to identify the target hole in training sessions and d latency time during memory phases were quantified in the Barnes maze. In the fear conditioning tests, freezing times were quantified e in context and f in response to a cue. All quantitative results (mean ± SD) shown were obtained with n = 8 animals per group. * P < 0.05 vs. control; # P < 0.05 vs. surgery. CXCL13 C-X-C motif chemokine 13, RT - qPCR reverse transcription-quantitative polymerase chain reaction, shRNA short hairpin RNA, sh - CRTL scrambled shRNA, sh - CXCL13 CXCL13 shRNA

Article Snippet: Membranes were blocked, then incubated overnight at 4 °C with antibodies against CXCL13 (1:100; Novus Biologicals, Littleton, CO, USA) or CXCR5 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA); or antibodies against ERK (1:500), p-ERK (1:500), IL-1β (1:1000), or TNF-α (1:500) (Cell Signaling, Beverly, MA, USA).

Techniques: Knockdown, shRNA, Western Blot, Quantitative RT-PCR, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Journal of Neuroinflammation

Article Title: Chemokine CXCL13 acts via CXCR5-ERK signaling in hippocampus to induce perioperative neurocognitive disorders in surgically treated mice

doi: 10.1186/s12974-020-02013-x

Figure Lengend Snippet: CXCL13 induces CXCR5/ERK-dependent release of pro-inflammatory cytokines in mouse hippocampus. WT or CXCR5 −/− mice received CXCL13 via intracerebroventricular injection. Seven days later, Barnes maze and fear conditioning tests were conducted. a Time to identify the target hole during training sessions and b latency time during memory phase were measured in the Barnes maze. Freezing times were quantified c in context and d in response to a cue. e Western blotting and densitometry of hippocampal p-ERK, total ERK, IL-1β, and TNF-α. Mice were injected with CXCL13 with or without PD98059 pretreatment, and behavioral tests were performed 7 days later. f Time to identify the target hole in training sessions and h latency time in memory phases of the Barnes maze. Freezing times were quantified i in context and j in response to a cue in fear conditioning tests. k Western blot and densitometry of IL-1β and TNF-α. All quantitative results (mean ± SD) shown were obtained with n = 8 animals per group. * P < 0.05 vs. vehicle + WT; # P < 0.05 vs. CXCL13 + WT; $ P < 0.05 vs. vehicle; & P < 0.05 vs. CXCL13. CXCL13 C-X-C motif chemokine 13, CXCR5 C-X-C motif chemokine receptor 5, WT wild-type, TNF - α tumor necrosis factor alpha, IL - 1β interleukin-1 beta

Article Snippet: Membranes were blocked, then incubated overnight at 4 °C with antibodies against CXCL13 (1:100; Novus Biologicals, Littleton, CO, USA) or CXCR5 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA); or antibodies against ERK (1:500), p-ERK (1:500), IL-1β (1:1000), or TNF-α (1:500) (Cell Signaling, Beverly, MA, USA).

Techniques: Injection, Western Blot