biotin conjugated isolectin b4  (Vector Laboratories)

Journal: bioRxiv

Article Title: Descending locus coeruleus noradrenergic signaling to spinal astrocyte subset is required for stress-induced pain facilitation

doi: 10.1101/2024.11.14.623627

Figure Lengend Snippet: (A and B) PWT before and 30 min after intrathecal administration of NA (0.1 nmol) in wild-type mice pretreated intrathecally with vehicle or CPT (3 nmol) (A: n = 5 mice per group) or in control ( Vgat-Cre mice with intra-SDH of AAV-flex[mCherry]) and SDH-Vgat + IN–A 1 R cKD mice (B: n = 9 mice per group) (two-way ANOVA with post hoc Bonferroni’s multiple comparisons test; *** P < 0.001). (C and D) PWT before and after acute restraint stress in wild-type mice pretreated intrathecally with vehicle or CPT (C: Vehicle, n = 6 mice; CPT, n = 5 mice) or in control ( Vgat-Cre mice with intra-SDH of AAV-flex[mCherry]) and SDH-Vgat + IN–A 1 R cKD mice (D: Control, n = 6 mice; SDH-Vgat + IN–A 1 R cKD, n = 7 mice) (two-way ANOVA with post hoc Bonferroni’s multiple comparisons test; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P <0.0001 vs. vehicle or control group). (E) Representative images of pERK (green) and IB4 (magenta) immunofluorescence in the SDH with or without Aβ fiber stimulation and/or restraint stress. CPT was intrathecally administered 30 min before stress exposure. (F) Quantitative analysis of the number of pERK + cells in superficial laminae of the SDH in each group ( n = 4–5 mice per group; one-way ANOVA with post hoc Tukey’s multiple comparisons test; * P < 0.05, *** P < 0.001). (G) Schematic illustration of the mechanisms of stress-induced pain facilitation highlighting NA signals from LC →SDH -NAergic terminals to Hes5 + astrocytes and Vgat + INs. Data represent mean ± SEM.

Article Snippet: Transverse spinal cord and brain sections (30 μm) were incubated in blocking solution (3% normal goat serum [#S-1000; Vector Laboratories] or normal donkey serum [#017-000-121; Jackson ImmunoResearch]) for 2 hours at room temperature and then incubated for 48 hours at 4°C with primary antibodies: polyclonal rabbit anti-tyrosine hydroxylase (TH; 1:1000; #AB152; Millipore); polyclonal sheep anti-TH (1:1000; #AB1542; Millipore); monoclonal mouse anti-noradrenaline transporter (NET; 1:2000; #NET05-2; Mab Technologies); polyclonal rabbit anti-green fluorescent protein (GFP; 1:1000; #598; MBL International); monoclonal rabbit anti-hemagglutinin (HA)-tag (1:1000; #3724; Cell Signaling); monoclonal rat anti-glial fibrillary acidic protein (GFAP; 1:2000; #13-0300; Invitrogen); polyclonal goat anti-SRY-related high-mobility group box 9 (SOX9; 1:1000; # AF3075; R&D Systems); polyclonal goat anti-paired box 2 (PAX2; 1:500; # AF3364; R&D Systems); monoclonal rat anti-mCherry (1:2000; #M11217; Thermo Fisher Scientific); polyclonal rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (pERK; 1:500; #9101; Cell Signaling); anti-isolectin B4 (IB4)-biotin conjugate (1:1000; # I21414; Thermo Fisher Scientific).

Techniques: Control, Immunofluorescence

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Fetal growth restriction and placental defects in obese mice are associated with impaired decidualisation: the role of increased leptin signalling modulators SOCS3 and PTPN2

doi: 10.1007/s00018-024-05336-7

Figure Lengend Snippet: Maternal obesity affects fetal-placental growth at embryonic day (E) 18.5. A Number of implantation sites at E6.5 and E18.5 pregnancies in chow-diet (CD) and high-fat diet (HFD) mice. B Representative images of fetuses at E18.5 collected from CD and HFD mice. C Analysis of fetal length and weight at E18.5 and D placental length and weight at E18.5 collected from CD and HFD mice, n indicates number of fetuses or placentas per diet. E Placental efficiency is measured by the fetal-weight-to-placental weight ratio index (FPI). F Haematoxylin and eosin (H&E) of midsagittal sections of E18.5 placentas from CD and HFD mice—labyrinthine zone (LZ) and junctional zone (JZ). G Isolectin BSI-B4 immunohistochemistry of fetal vasculature in the placental labyrinth at E18.5 in placentas collected from CD and HFD mice. H Ratio labyrinth and junctional zone to the placental area at E18.5 in placentas collected from CD and HFD mice, n indicates number of placental sections from 3 to 4 placentas per mother/diet, n = 3 CD and n = 4 HFD. All data are mean ± SEM with individual values from placental sections. Statistical analysis between groups was carried out using Mann–Whitney. *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: The primary antibodies used were: anti-KI67 (ab15580; Abcam, Cambridge, United Kingdom), anti-PR (D8Q2J; Cell Signaling Technology, Danvers, USA), anti-SOCS3 (ab16030; Abcam), anti-PTPN2 (ab180764; Abcam), anti-pSTAT3 (ab86430; Abcam), Anti-Vimentin (D21H3; Cell Signaling Technology), anti-pan Cytokeratin (C2562, Sigma Aldrich, Saint Louise, USA), anti-CDH1 (610,181, BD Biosciences, New Jersey) and biotin-conjugated isolectin from Bandeiraea simplicifolia BSI-B4 (L2140, Sigma).

Techniques: Immunohistochemistry, MANN-WHITNEY