Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: Identification of DEGs involved in the pathogenesis of CPAM. (a) Flowchart of the study design and samples at each stage of analysis. (b) Scatterplot of mRNA expression variation between diseased CPAM and normal tissues. (c) Hierarchical cluster of gene expression profiles from microarray assays. Expression of CA12, LONRF3, MAP2, THBS1, and PPID at mRNA (d) and protein (e) levels in lung tissues from CPAM and adjacent normal tissues. ∗∗∗ p < 0.001, compared with normal.

Article Snippet: Primary antibodies for this study were specific for CA12 (Boster, Cat. No. A04063, 1 : 1000), LONRF3 (GeneTex, Cat. No. GTX112150, 1 : 2000), MAP2 (Boster, Cat. No. A01201, 1 : 2000), THBS1 (Boster, Cat. No. PB0471, 1 : 2000), PPID, E-cadherin (Beyotime, Cat. No. AF6759, 1 : 1000), N-cadherin (Beyotime, Cat. No. AF5237, 1 : 800), aggrecan (Abcam, Cat. No. ab3778, 1 : 1000), Col2A1 (Boster, Cat. No. A00517, 1 : 2000), MMP13 (Proteintech, Cat. No. 18165-1-AP, 1 : 3000), ADAMTS4 (Proteintech, Cat. No. 11865-1-AP, 1 : 600), and GAPDH (Abcam, Cat. No. ab9485, 1 : 2000).

Techniques: Expressing, Gene Expression, Microarray

Journal: Neuron

Article Title: Targeted proteoform mapping uncovers specific Neurexin-3 variants required for dendritic inhibition

doi: 10.1016/j.neuron.2022.04.017

Figure Lengend Snippet:

Article Snippet: The following antibodies were used in this study: rabbit polyclonal anti-β-actin (Abcam; Cat# ab8227; RRID: AB_2305186; LOT# GR3314266-1), mouse monoclonal anti-calbindin (Swant; Cat# 300; RRID: AB_10000347; LOT# 17 (F)), goat polyclonal anti-calretinin (Swant; Cat# CG1; RRID: AB_10000342; LOT# 1§.1), mouse monoclonal anti-CamKII alpha (Thermo Fisher Scientific; 6G9; Cat# Ma1-048; RRID: AB_325403; LOT# TH269517), mouse monoclonal anti-cannabinoid receptor 1 (Immunogene; IMG-3C2; Cat# IMG-CB1R-mAb001; LOT# CJ03), guinea pig polyclonal anti-cholecystokinin (Synaptic Systems, Cat# 438004; RRID: AB_2814938; LOT# 1-1), mouse monoclonal anti-GAD67 (Millipore; 1G10.2; Cat# MAB5406; RRID: AB_2278725; LOT# 3015328), rabbit polyclonal anti-GAPDH (Enogene; Cat# E1C604; LOT# R14Q12), mouse monoclonal anti-gephyrin (Synaptic Systems; mAb7a; Cat# 147021; RRID: AB_2232546; LOT# 147021/15), mouse monoclonal anti-gephyrin (Synaptic Systems; mAb7a; Cat# 147011; RRID: AB_887717; LOT# 147011/54), rat monoclonal anti-HA (Roche; 3F10; Cat# 11867431001; RRID: AB_390919; LOT# 34502100), rabbit monoclonal anti-HA (Cell Signaling; 3724; Cat# 3724; RRID: AB_1549585; LOT# 9), guinea pig polyclonal anti-MAP2 (Synaptic Systems; Cat# 188004; RRID: AB_2138181; LOT# 2-26), mouse monoclonal anti-MAP2 (Synaptic Systems; 198A5; Cat# 188011; RRID: AB_2147096; LOT# 1-10), rabbit polyclonal anti-neurexin , chicken anti-neurexin , rabbit anti-neuroligin , rabbit monoclonal anti-nNOS (Cell Signaling; C7D7; Cat# 4231; RRID: AB_2152485; LOT# 2), goat polyclonal anti-parvalbumin (Swant, Cat# PVG214; RRID: AB_10000345), mouse monoclonal anti-PSD95 (Santa Cruz; 7E3; Cat# sc32290; RRID: AB_628114; LOT# J1509), goat polyclonal anti-somatostatin (Santa Cruz; Cat# sc7819; RRID: AB_2302603; LOT# L1611), mouse monoclonal anti-Synaptotagmin 2 (Zebrafish International Resource Center; Cat# znp-1; RRID: AB_10013783), mouse monoclonal anti-V5 (Biorad; SV5-PK1; Cat# MCA1360; RRID: AB_322378; LOT# 148239), guinea pig polyclonal anti-vGAT (Synaptic Systems; Cat# 131004; RRID: AB_887873; LOT# 2-42), guinea pig polyclonal anti-vGlut1 (Millipore; Cat# AB5905; RRID: AB_2301751; LOT# 3308226), rabbit polyclonal anti-VIP (Immunostar; Cat# 20077; RRID: AB_572270; LOT# 1513001), guinea pig anti-somatostatin (raised in the present study against amino acid residues 35-88 of mouse pro-somatostatin, GenBank: #BC010770.1), goat anti-cannabinoid receptor 1 (Nittobo Medical, MSFR100600; RRID: AB_2571592), guinea pig anti-GABAA receptor alpha 1 (Nittobo Medical, MSFR101540; RRID: AB_2571572), goat anti-vGAT (Nittobo Medical, MSFR106130; RRID: AB_2571623), guinea pig anti-PSD95 (Nittobo Medical, MSFR105180; RRID: AB_2571612).

Techniques: Recombinant, Reporter Assay, Multiplex Assay, Plasmid Preparation, Software, Microscopy

Journal: Molecular Psychiatry

Article Title: A CRHR1 antagonist prevents synaptic loss and memory deficits in a trauma-induced delirium-like syndrome

doi: 10.1038/s41380-020-0659-y

Figure Lengend Snippet: a IHC for Shank2 (green), Vglut1 (red), MAP2 (magenta) (scale bar = 5 µm), western blot analysis for all different treatment regimens (CRH; the CRHR1 blocker NBI30775; the corticosterone receptor blocker RU38486; and BDNF) and the relative quantification. b IHC for Shank2 (green), Vglut1 (red), MAP2 (magenta) and CRHR1 (white) with colocalization and trilocalization analysis (scale bar = 5 µm). c IHC for phospho-NF-κB p65 (green, scale bar = 5 μm) and the relative quantification of the signal within the nuclear compartment. d IHC for phospho-IKB-α and the relative quantification of the signal within the cytosolic compartment. e IHC for and compartment analysis of p65. f IHC (left panel) for Shank2 (green), Vglut1 (red), MAP2 (magenta) (scale bar = 5 µm), WB of BDNF (right panel) for all different treatment regimens blocking the NF-κB pathway (JSH (the translocation blocker JSH- 23); SC (the IKK-β inhibitor SC-514); and LAC (the NF-κB activation blocker lactacystin)) and the relative quantification. g IHC for ATG5, immunoblot analysis of h BCN-1, i LC3, and j Lamp-2 and the relative quantification normalized to β-actin. k TEM of acquired synapses after 15 min of CRH treatment (arrows indicate membrane-associated vesicles (scale bar = 0.5 µm)) and the relative quantification of docked vesicles and multivesicular bodies (MVBs)/synapses (a two-tailed unpaired T -test was used). l Quantification of the number of excitatory synapses (left) by IHC and WB analysis of BDNF expression (right) after treatment with 100 nM CRH for 30 min and Leupeptin hemisulfate + 5 μM E64 (CRH++) compared with control treatment. N = 3–5; the error bars represent the SEMs; one-way ANOVA and Bonferroni’s post hoc comparison test were performed (* P ≤ 0.05, ** P ≤ 0.005, *** P ≤ 0.0005, **** P ≤ 0.0001).

Article Snippet: The following primary antibodies were purchased from commercial suppliers: Vglut1 (1:500, Synaptic Systems GmbH, #135304), Gephyrin (1:500, Synaptic Systems GmbH, #147003), Vgat (1:500, Synaptic Systems GmbH, #131011), Iba1 (1:250, Wako Chemical GmbH, #NCNP24), GFAP (1:500, Sigma-Aldrich, #G3893 ) , NeuN (1:1000, Millipore, #MAB377), C1q (1:1000, Abcam, #182451), CRH (1:1000, Abcam, #8901), p62 (1:500, Abcam, #56416), Map2 (1:500, EnCor Biotechnology Inc., #CPCA-MAP2), Synaptotagmin-1 (1:500, Synaptic Systems, #105311C5), ATG5 (Abcam, #AB108327), NF-κB p65 (1:500, Santa Cruz, #SC8008), phospho-NF-κB (Thr435) (1:500, Thermo Fisher Scientific, #PA5–37724), phospho-IκBα (Ser32) (1:500, Cell Signaling, #2859), CRHR1 (1:500, Everest Biotech, #EB08035), and ChAT (1:250, Synaptic System, #297015).

Techniques: Western Blot, Blocking Assay, Translocation Assay, Activation Assay, Two Tailed Test, Expressing