l pgds antibody  (Proteintech)

Journal: Cells

Article Title: L-PGDS–PGD2–DP1 Axis Regulates Phagocytosis by CD36 + MGs/MΦs That Are Exclusively Present Within Ischemic Areas After Stroke

doi: 10.3390/cells13201737

Figure Lengend Snippet: ( A ) L-PGDS expression by WB from samples collected from MCA areas of sham-operated mice or ischemic areas at 1, 3, 5, and 7 days post-stroke. ( B , C ) WB analysis at 7 days post-stroke. L-PGDS expression levels were significantly higher in ischemic areas compared to the contralateral side of nonischemic areas. ( D – G ) Immunohistochemistry for L-PGDS and GFAP. L-PGDS was strongly detected in the leptomeninges of ischemic areas (L-PGDS [( E – G ): red]); GFAP [( E – G ): green]; DAPI [( E – G ): blue]). ( H – K ) Immunohistochemistry for L-PGDS and PDGFRβ. L-PGDS was co-expressed in PDGFRβ + pericytes in the leptomeninges of ischemic areas (( I – K ), arrows) (L-PGDS [( I , J ): red]; PDGFRβ [( I , K ): green]; DAPI [( I – K ): blue]). ( L – N ) RT-qPCR analysis. L-PGDS levels were significantly higher in brain pericytes treated with supernatant from ischemic areas ( M , N ) than in untreated pericytes (control) ( L , N ). Scale bars: ( E – G ) 100 µm; ( I ) 20 µm. * p < 0.05 between the contralateral side of nonischemic areas and ischemic areas, n = 3 for each area ( C ). * p < 0.05 between the control and supernatant–treated group, n = 3 for each group ( N ). Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; GFAP, glial fibrillary acidic protein; L-PGDS, lipocalin-type prostaglandin D synthase; MCA, middle cerebral artery; PDGFRβ, platelet-derived growth factor receptor-β; RT-qPCR, quantitative reverse transcription-polymerase chain reaction; WB, Western blot.

Article Snippet: The membranes were incubated with primary antibodies against L-PGDS (1:1000, mouse; Santa Cruz Biotechnology), CD36 (1:2000, goat; R&D Systems), and β-actin (1:2000, mouse; Sigma-Aldrich, St. Louis, MO, USA), followed by incubation with horseradish peroxidase-labeled secondary antibodies (1:2000, mouse; Cell Signaling Technology, Danvers, MA, USA, and 1:1000, goat; Santa Cruz Biotechnology).

Techniques: Expressing, Immunohistochemistry, Quantitative RT-PCR, Control, Derivative Assay, Reverse Transcription, Polymerase Chain Reaction, Western Blot

Journal: eLife

Article Title: Oligodendrocyte-lineage cell exocytosis and L-type prostaglandin D synthase promote oligodendrocyte development and myelination

doi: 10.7554/elife.77441

Figure Lengend Snippet: Figure 7. The effect of the L-PGDS inhibitor, AT-56, and HPGD, which inactivates PGD2, on oligodendrocyte development in vitro. (A) Immunoblot of secreted and intracellular L-PGDS protein from oligodendrocyte cultures from PD:ibot and littermate control mice. (B) Quantification of the immunoblot signal intensity of secreted L-PGDS. N=10 mice per group. Paired two-tailed T-test. Secreted L-PGDS intensity: 3.0±0.4 in control and 2.2±0.4 in PD:ibot, p=0.0104. (C) Quantification of the ratio of secreted and intracellular L-PGDS. N=10 mice per group. Paired two-tailed T-test. Ratio of secreted and intracellular L-PGDS: 37.2±10.4 in control and 18.9±3.7 in PD:ibot, p=0.05. (D) Oligodendrocyte cultures from wild-type mice after 7 days of differentiation in the presence and absence of the L-PGDS inhibitor AT-56. Red: MBP immunofluorescence. Blue: CellMask, which labels all cells. Scale bars: 50 μm. (E) Quantification of cells with lamellar morphology. One-way ANOVA with Benjamini, Krieger, and Yekutieli’s two-stage linear step-up FDR procedure for multiple comparisons. N=3 cultures from 3 mice per group. Lamellar cells%: DMSO control: 28±0.8; 1 μM AT-56: 15±1.6, p=0.024; 5 μM AT-56: 7.3±1.3, p=0.0068. (F) Oligodendrocyte cultures from wild-type mice after 7 days of differentiation in the presence and absence of HPGD. Green: Membrane version of CellMask, which labels the membranes of all cells. Blue: DAPI. Scale bars: 50 μm. (G) Quantification of cells with lamellar morphology. One-way ANOVA with Benjamini, Krieger, and Yekutieli’s two-stage linear step-up FDR procedure for multiple comparisons. N=7 cultures from 6 mice per group. Lamellar cells%: 0 μM HPGD: 59.10±5.33; 3 μM HPGD: 50.09±5.80, p=0.034; 6 μM HPGD: 42.20±5.87, p=0.0090.

Article Snippet: Membranes were blocked with clear milk blocking buffer (Fisher, cat #PI37587) for 1 hr at room temperature and incubated with primary antibodies against L- PGDS (Santa Cruz Biotechnology, cat #sc- 390717, dilution 1:1000), GAPDH (Sigma, cat #CB1001, dilution 1:5000), BoNT- B Light Chain (R&D Systems, cat #AF5420- SP, dilution 1:1000), VAMP2 (Synaptic Systems, cat #104 211, dilution 1:1000), VAMP3 (Novus Biological, cat # NB300- 510- 0.025mg, dilution 1:1000) and MBP (Abcam, cat #ab7349, dilution 1:1000) at 4 °C overnight.

Techniques: In Vitro, Western Blot, Control, Two Tailed Test, Immunofluorescence, Membrane

Journal: eLife

Article Title: Oligodendrocyte-lineage cell exocytosis and L-type prostaglandin D synthase promote oligodendrocyte development and myelination

doi: 10.7554/elife.77441

Figure Lengend Snippet: Figure 8. Oligodendrocyte defect in L-PGDS-knockout mice. (A) CC1 immunofluorescence at P9. The dashed lines delineate the corpus callosum (CC). Ctx, cortex. Str, striatum. Scale bar: 200 μm. (B) Quantification of the density of CC1+ cells. N=4 mice per genotype. Corpus callosum: 217.6±20.3 / mm2 in control, 114±22.8 in knockout, p=0.015; cerebral cortex: 42.5±2.2 in control, 14.3±2.0 in knockout, p=0.0001. Unpaired two-tailed T-test in all quantifications in this figure. (C) PDGFRα immunofluorescence at P9 in the corpus callosum and the cerebral cortex. Scale bar: 20 μm. (D) Quantification

Article Snippet: Membranes were blocked with clear milk blocking buffer (Fisher, cat #PI37587) for 1 hr at room temperature and incubated with primary antibodies against L- PGDS (Santa Cruz Biotechnology, cat #sc- 390717, dilution 1:1000), GAPDH (Sigma, cat #CB1001, dilution 1:5000), BoNT- B Light Chain (R&D Systems, cat #AF5420- SP, dilution 1:1000), VAMP2 (Synaptic Systems, cat #104 211, dilution 1:1000), VAMP3 (Novus Biological, cat # NB300- 510- 0.025mg, dilution 1:1000) and MBP (Abcam, cat #ab7349, dilution 1:1000) at 4 °C overnight.

Techniques: Knock-Out, Immunofluorescence, Control, Two Tailed Test

Journal: eLife

Article Title: Oligodendrocyte-lineage cell exocytosis and L-type prostaglandin D synthase promote oligodendrocyte development and myelination

doi: 10.7554/elife.77441

Figure Lengend Snippet: Figure 9. Rescue of oligodendrocyte deficit in PD:ibot by L-PGDS protein, PGD2 in vitro, and myelination deficit in PD:ibot by L-PGDS overexpressing transgenic mice. (A) Oligodendrocyte cultures from wild-type mice after 7 days of differentiation in the presence and absence of L-PGDS protein. Green: Membrane-staining version of CellMask, which labels the membranes of all cells. Blue: DAPI. Scale bars: 50 μm. (B) Quantification of cells with lamellar morphology. One-way ANOVA with Benjamini, Krieger, and Yekutieli’s two-stage linear step-up FDR procedure for multiple comparisons.

Article Snippet: Membranes were blocked with clear milk blocking buffer (Fisher, cat #PI37587) for 1 hr at room temperature and incubated with primary antibodies against L- PGDS (Santa Cruz Biotechnology, cat #sc- 390717, dilution 1:1000), GAPDH (Sigma, cat #CB1001, dilution 1:5000), BoNT- B Light Chain (R&D Systems, cat #AF5420- SP, dilution 1:1000), VAMP2 (Synaptic Systems, cat #104 211, dilution 1:1000), VAMP3 (Novus Biological, cat # NB300- 510- 0.025mg, dilution 1:1000) and MBP (Abcam, cat #ab7349, dilution 1:1000) at 4 °C overnight.

Techniques: In Vitro, Transgenic Assay, Membrane, Staining

Journal: International Journal of Molecular Sciences

Article Title: Indomethacin Disrupts the Formation of β-Amyloid Plaques via an α2-Macroglobulin-Activating lrp1-Dependent Mechanism

doi: 10.3390/ijms22158185

Figure Lengend Snippet: Indomethacin treatment suppresses the expression of COX-2, L-PGDS and CRTH2 proteins. ( A , B ) APP/PS1 Tg mice were intragastrically administered the indicated concentrations of indomethacin ranging from 0.01 to 5 mg/kg. Western blotting was employed to detect the levels of COX-2, L-PGDS and CRTH2. β-actin served as an internal control. ( C ) N2a cells were treated with the indicated concentrations of indomethacin ranging from 10 to 500 μM for 24 h. Western blotting was used to determine the expression of COX-2, L-PGDS and CRTH2. β-actin served as an internal control. The optical density of the bands was analyzed using ImageJ software (v1.51, NIH, Bethesda, MD, USA). Data are presented as means ± S.E. of independent experiments. * p < 0.05; ** p < 0.01 and *** p < 0.001 compared to vehicle-treated controls.

Article Snippet: Antibodies specific for A2M, L-PGDS and CRTH2 were purchased from Santa Cruz Biotechnology (Shanghai, China).

Techniques: Expressing, Western Blot, Software

Journal: International Journal of Molecular Sciences

Article Title: Indomethacin Disrupts the Formation of β-Amyloid Plaques via an α2-Macroglobulin-Activating lrp1-Dependent Mechanism

doi: 10.3390/ijms22158185

Figure Lengend Snippet: PGD 2 suppresses the expression of the A2M protein in a CRTH2-dependent manner. ( A ) APP/PS1 Tg mice were injected (i.c.v.) with the indicated doses of PGD 2 ranging from 0.1 to 2 μg. After 24 h, Western blotting was used to determine the expression of the A2M protein. β-actin served as an internal control. ( B ) N2a cells were treated with the indicated concentration of PGD 2 from 1 to 20 μM for 24 h. ( C ) In selected experiments, N2a cells were treated with 10 μM PGD 2 in the absence or presence of the indicated concentrations of a CRTH-2 inhibitor, Bay-u3405, ranging from 1 to 10 μM for 24 h. Western blotting was used to determine the expression of A2M. β-actin served as an internal control. The optical density of the bands was analyzed using ImageJ software (v1.51, NIH, Bethesda, MD, USA). Data are presented as the means ± S.E. of independent experiments. * p < 0.05; ** p < 0.01 and *** p < 0.001 compared to vehicle-treated controls. ## p < 0.01 compared to PGD 2 -treated groups.

Article Snippet: Antibodies specific for A2M, L-PGDS and CRTH2 were purchased from Santa Cruz Biotechnology (Shanghai, China).

Techniques: Expressing, Injection, Western Blot, Concentration Assay, Software