goat polyclonal anti olig2  (R&D Systems)

Journal: bioRxiv

Article Title: Integrated 5-HT 2A –TrkB and G protein signaling in serotonergic psychedelic responses

doi: 10.64898/2026.03.19.712961

Figure Lengend Snippet: a. Schematic representation of the assay used to quantify synaptic connectivity by rabies virus neuronal tracing. Rabies virus particles (in red), encoding the mCherry fluorescent reporter and pseudotyped with the EnvA envelope glycoprotein, selectively infect NSC TVA derived neurons (in green) that express the TVA receptor and EGFP. The virus is then transmitted retrogradely to presynaptic NSC shRNA TrkB or NSC shRNA 5-HT 2A derived neurons (in pink). Synaptic inputs are quantified by counting mCherry-positive (red) cells using image analysis. b. Undifferentiated neural stem cells (NSCs) expressing TVA receptors, EGFP, and the rabies glycoprotein were selected by flow cytometry based on EGFP fluorescence. Cells were then infected with different dilutions of EnvA-pseudotyped rabies virus expressing mCherry, which appears as red fluorescence in the lower panels. Images were acquired at 10x magnification. c. Representative microscopic fields of neurons derived from NSC TVA differentiation, exhibiting mCherry fluorescence 48 hours post-rabies virus infection. Scale bar: 20 µm. d. Evaluation of the proportion of NSC TVA cells seeded in mixture with wild type NSCs to achieve an optimal ratio of total mCherry expressing neurons relative to EGFP expressing neurons. Left axis: ratio of mCherry+ to EGFP+ cells. Right axis: percentage of EGFP+ cells. e. Evaluation of the proportion of NSC TVA cells seeded in mixture with wild type NSCs to identify an optimal window for quantifying the effects of neuroplastogenic treatments on mCherry+ neuron numbers. Cells were treated with BDNF (50 ng/mL) for 24 h, and synaptogenesis was assessed according to the established protocol (see Methods). Data were analyzed by two-way ANOVA, revealing significant effects of cell proportion (F(3,71) = 280.1, P < 0.0001), treatment (F(1,71) = 48.03, P < 0.0001), and their interaction (F(3,71) = 23.43, P < 0.0001). ****p < 0.0001.

Article Snippet: The membranes were then incubated overnight with either rabbit polyclonal anti-5-HT 2A antibody (1:500, Immunostar #242800) or goat polyclonal anti-TrkB antibody (1:1,000; R&D Systems AF1494).

Techniques: Virus, Derivative Assay, shRNA, Expressing, Flow Cytometry, Fluorescence, Infection

Journal: bioRxiv

Article Title: Integrated 5-HT 2A –TrkB and G protein signaling in serotonergic psychedelic responses

doi: 10.64898/2026.03.19.712961

Figure Lengend Snippet: a. Fluorescence microscopy images from immunocytochemistry assays showing the distribution of TrkB receptors (left column) in neural cells differentiated from neural stem cells. The upper row demonstrates co-expression of TrkB receptors with the neuronal marker MAP2, whereas the middle row shows the presence of TrkB receptors in cells expressing the glial marker GFAP. The lower row shows co-expression of TrkB and 5-HT 2A receptors in both neurons and glial cells. Scale bar: 20 µm. b. Immunoblots displaying protein bands corresponding to TrkB receptors (top) and 5-HT 2A receptors (bottom). Lane 1 shows undifferentiated neural stem cells and lane 2 shows neural cells differentiated for 5 days. c. Effect of IPTG treatment on the expression of Ntrk2 and Htr2a mRNAs in non-differentiated NSC shRNA 5-HT 2A , differentiated NSC shRNA 5-HT 2A , non-differentiated NSC shRNA TrkB and differentiated NSC shRNA TrkB, as determined by RT-qPCR. d. Immunoblots showing the effect of IPTG treatment on the expression of TrkB receptors (top) and 5-HT 2A receptors (bottom) in cell lysates from NSC shRNA TrkB and NSC shRNA 5-HT 2A lines differentiated for 5 days. Immunoblots showing 40 KDa bands correspond to loading controls made with actin immunoreactivity.

Article Snippet: The membranes were then incubated overnight with either rabbit polyclonal anti-5-HT 2A antibody (1:500, Immunostar #242800) or goat polyclonal anti-TrkB antibody (1:1,000; R&D Systems AF1494).

Techniques: Fluorescence, Microscopy, Immunocytochemistry, Expressing, Marker, Western Blot, shRNA, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Integrated 5-HT 2A –TrkB and G protein signaling in serotonergic psychedelic responses

doi: 10.64898/2026.03.19.712961

Figure Lengend Snippet: a. Wild-type NSCs (grey bars), TrkB-silenced NSCs (NSC shRNA TrkB, yellow bars), and 5-HT 2A -silenced NSCs (NSC shRNA 5-HT 2A , red bars) were differentiated into neurons and glia and treated with the indicated drugs. Bars represent the fold change of the area under the curve (AUC) obtained from Sholl analysis. Two-way ANOVA revealed significant main effects of phenotype (F(2,60) = 226.600, p < 0.0001) and treatment (F(9,60) = 9.614, p < 0.0001), as well as a significant interaction (F(18,60) = 5.620, p < 0.0001). Post hoc comparisons were performed using Tukey’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. # indicates significant differences between WT and NSC shRNA TrkB. Same p values are applied. b. Cell mixtures of NSC TVA with wild- type NSCs (grey bars), TrkB-silenced NSCs (NSC shRNA TrkB, yellow bars), and 5-HT 2A -silenced NSCs (NSC shRNA 5-HT 2A , red bars) were differentiated into neurons and glia and treated with the indicated drugs. Bars represent the relative number of mCherry-positive cells versus vehicle. Statistical analysis was performed using two-way ANOVA, which revealed significant effects of phenotype (F(2,72) = 2008, p < 0.0001), drug treatment (F(11,72) = 23.44, p < 0.0001), and their interaction (F(22,72) = 17.81, p < 0.0001). Post hoc comparisons were performed using Tukey’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. # indicates significant differences between the silenced condition and WT among applying vehicles.

Article Snippet: The membranes were then incubated overnight with either rabbit polyclonal anti-5-HT 2A antibody (1:500, Immunostar #242800) or goat polyclonal anti-TrkB antibody (1:1,000; R&D Systems AF1494).

Techniques: shRNA, Comparison

Journal: bioRxiv

Article Title: Integrated 5-HT 2A –TrkB and G protein signaling in serotonergic psychedelic responses

doi: 10.64898/2026.03.19.712961

Figure Lengend Snippet: Graphs illustrate a representative neuroplasticity experiment showing Sholl curves from neurons derived from NSC shTrkB or NSC sh5-HT 2A cell lines treated with the indicated compounds, compared with their respective vehicle controls. Columns designated as IPTG+ correspond to cells pretreated with IPTG to silence the expression of either TrkB or 5-HT 2A receptors.

Article Snippet: The membranes were then incubated overnight with either rabbit polyclonal anti-5-HT 2A antibody (1:500, Immunostar #242800) or goat polyclonal anti-TrkB antibody (1:1,000; R&D Systems AF1494).

Techniques: Derivative Assay, Expressing

Journal: bioRxiv

Article Title: Integrated 5-HT 2A –TrkB and G protein signaling in serotonergic psychedelic responses

doi: 10.64898/2026.03.19.712961

Figure Lengend Snippet: Wild-type NSCs (grey bars), TrkB-silenced NSCs (NSC shRNA TrkB, yellow bars), and 5-HT 2A -silenced NSCs (NSC shRNA 5-HT 2A , red bars) were differentiated into neurons and glia and treated with the indicated drugs to assess changes in immediate early gene expression. a. c-Fos expression, represented as log2 fold change relative to vehicle levels. Two-way ANOVA revealed significant effects of drug treatment (F(10,66) = 7.214, p < 0.0001), NSC phenotype (F(2,66) = 50.07, p < 0.0001), and their interaction (F(20,66) = 3.231, p = 0.0002). Post hoc comparisons were performed using Tukey’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. b. Egr-2 expression, represented as log2 fold change relative to vehicle levels. Two-way ANOVA revealed significant effects of drug treatment (F(10,66) = 6.872, p < 0.0001), NSC phenotype (F(2,66) = 85.90, p < 0.0001), and their interaction (F(20,66) = 3.162, p = 0.0002). Post hoc comparisons were performed using Tukey’s multiple comparison test versus vehicle. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The membranes were then incubated overnight with either rabbit polyclonal anti-5-HT 2A antibody (1:500, Immunostar #242800) or goat polyclonal anti-TrkB antibody (1:1,000; R&D Systems AF1494).

Techniques: shRNA, Gene Expression, Expressing, Comparison

Journal: STAR Protocols

Article Title: Protocol to study adult neurogenesis in fresh-frozen human hippocampal tissue using an immunofluorescence quantitative approach

doi: 10.1016/j.xpro.2025.104344

Figure Lengend Snippet: Overview of expected outcomes (A) Schematic representation of distinct cell populations related to adult hippocampal neurogenesis (AHN). (B–H) Expected staining with anti- (B) vimentin, (C) sex determining region Y-box 2 (Sox2), (D) phospho-histone 3 (PH3), (E) HuC-HuD, (F) Doublecortin (DCX), (G) polysialylated-neural cell adhesion molecule (PSA-NCAM), and (H) calbindin (CB) antibodies. (I) Schematic representation of the components of the human hippocampal neurogenic niche. (J–M) Expected staining with anti- (J) S100 calcium-binding protein β (S100β), (K) ionized calcium-binding adaptor molecule 1 (Iba1), (L) phosphorylated γH2A.X (γH2A.X) antibodies, and (M) Ulex Europaeus Agglutinin-I (UEA-1). Yellow scale bar: 20 μm. White scale bar: 10 μm. Yellow triangles indicate positive cells. White triangles highlight the morphology of cells labelled for each marker.

Article Snippet: Goat polyclonal anti-SRY (sex-determining region Y)-box 2 (Sox2) (1:500) , R and D Systems , Cat# AF2018; RRID: AB_355110.

Techniques: Staining, Binding Assay, Marker