rabbit anti ctip2 bcl 11b  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Imp1 acts as a dosage- and stage-dependent temporal rheostat orchestrating radial glial fate transitions and cortical morphogenesis

doi: 10.1101/2025.11.18.688993

Figure Lengend Snippet: (A,D,G) Representative images of control, continuous Imp1 overexpression, and T1 Imp1 conditions showing TEMPO reporters, Cux1/Ctip2 immunostaining and overlays. Boxed regions: Ctip2+ TEMPO neurons (dashed) or double-positive Cux1+/Ctip2+ TEMPO cells (solid). (B,E,H) High magnification images of boxed regions highlight CFP-/RFP-labeled neurons in layers V-VI colocalizing with Ctip2 (outlined arrowheads) or double-positive for both markers (solid arrowheads). (C,F,I) Quantification of marker expression in CFP+ and RFP+ neurons residing in layers V-VI. Following continuous or T1 Imp1 overexpression, neurons in deep layer maintain appropriate deep-layer molecular identities (predominantly Ctip2+), demonstrating that laminar distribution reflects bona fide fate specification changes rather than mislocalization. (C) In control conditions, CFP+: Ctip2 (78.31% ± 13.82%), Cux1 (2.93% ± 2.11%), double-negative (18.76% ± 11.73%). RFP+: Ctip2 (50.35% ± 17.01%), Cux1 (1.85% ± 1.85%), double-negative (46.76% ± 17.20%), double-positive (1.04% ± 1.04%). (F) Following continuous Imp1 overexpression, CFP+: Ctip2+ (71.18% ± 3.06%), Cux1+ (7.38% ± 4.93%), double-negative (16.06% ± 1.93%), double-positive (5.38% ± 1.80%). RFP+: Ctip2+ (58.92% ± 7.18%), Cux1+ (5.84% ± 3.01%), double-negative (5.10% ± 3.12%), double-positive (30.14% ± 11.98%). (I) In T1 Imp1 overexpression, CFP+: Ctip2+ (47.26% ± 4.88%), Cux1+ (14.63% ± 3.76%), double-negative (36.41% ± 5.64%), double-positive (1.68% ± 0.57%). RFP+: Ctip2+ (47.38% ± 13.91%), Cux1+ (9.17% ± 4.68%), double-negative (26.86% ± 5.03%), double-positive (16.59% ± 10.02%). Dashed lines: upper (II-IV), lower cortical layers (V-VI) and subplate zone (SPZ). Scale bars: (A,D,G) 100 µm and (B,E,H) 20 µm. Data show mean±SEM. Statistics: two-tailed unpaired Welch’s t-test (*P < 0.05, **P < 0.01, *** P < 0.001).

Article Snippet: Primary antibodies were applied overnight to amplify TEMPO signals or detect fate markers at 4°C: mouse anti-V5 (1:650, Thermo Fisher R96025) to detect CFP+ cells, rat anti-mCherry (1:500, Thermo Fisher M11217) or chicken anti-mCherry (1:500, AvesLabs MCHERRY AB_2910557) to detect RFP+ cells, rat anti-Ctip2 (1:100, Abcam [25B6] ab18465) to detect Ctip2+ cells, and rabbit anti-Cux1 (1:50, Proteintech 11733-1-AP) to detect Cux1+ cells.

Techniques: Control, Over Expression, Immunostaining, Labeling, Marker, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Dynamic regulation of NeuroD1 expression level by a novel viral construct during astrocyte-to-neuron reprogramming

doi: 10.1101/2025.02.17.638625

Figure Lengend Snippet: The reprogrammed neurons by the ND1-124T-GFP construct can express markers of mature neurons and neuronal subtypes in long term cultures. ( A ) A live image of HA cell cultures infected by ND1-GFP retrovirus showing floating cell spheroids in the hanging drops of culture medium at 3 DPI. ( B ) Live images of the resulting cell spheroids that are attached to a monolayer of HA cells at 20 DPI. #, the core of the attached spheroid; Arrowhead, a HA cell infected by ND1-GFP that has migrated to periphery of the spheroid showing a typical neuronal morphology. At 30 DPI, the attached HA spheroid cultures infected by either ND1-GFP or ND1-124T-GFP were fixed and immunostained with the indicated mature neuronal markers (NeuN and/or Map2) along with NeuroD1 ( C ), the synaptic vesicle marker SV2 ( D ), the glutamatergic neuronal markers vGlut1 and HuD ( E ), and Ctip2 ( F ). ( G ) Expression levels of the markers were quantified by their cellular fluorescence intensities and compared. ( H ) The GABAergic interneuron markers GABA and GAD67 ( F ) were also analyzed and compared ( I ). Arrows, the marker+ cells. ns, not significant; ***, p < 0.005; ****, p < 0.001 by student t -test. Scale bars, 40μm.

Article Snippet: Briefly, fixed cell cultures were incubated with monoclonal antibodies against NeuroD1 (mouse IgG, 1:1000, Abnova), GFP (rat IgG2a, 1:400, BioLegend), HuD/ELAVL4 (mouse IgG, 1:200, Santa Cruz), GAD67 (mouse IgG, 1:200, Millipore), and NeuN (mouse IgG, 1:400, Millipore); polyclonal antibodies against GFP (chicken IgY, 1:400, Aves), mCherry/RFP (rabbit IgG, 1:500, Abcam), mCherry/RFP (chicken IgY, 1:400, Aves), Map2 (rabbit IgG, 1:400, Abcam), DCX (rabbit IgG, 1:500, Abcam), vGlut1 (guinea pig IgG, 1:200, Millipore), Ctip2 (guinea pig IgG, 1:200, Synapse Systems), anti-SV2 (rabbit, 1:2000, Developmental Studies Hybridoma Bank), GABA (rabbit IgG, 1:200, Calbiochem), and DCX (guinea pig IgG, 1:1000, Millipore), followed by appropriate species-specific secondary antibodies (Molecular Probes).

Techniques: Construct, Infection, Marker, Expressing, Fluorescence

Journal: bioRxiv

Article Title: Porphyromonas gingivalis outer membrane vesicles alter neuronal architecture and Tau phosphorylation in the embryonic mouse brain

doi: 10.1101/2024.09.03.611094

Figure Lengend Snippet: (A) Representative immunofluorescent images from sagittal sections of PFA fixed brains at GA 18.5 pup brains showing location of cortical layer markers Cux1, SatB2, and Ctip2. Approximate position of cortical layers 1-6 with the intraventricular zone (IZ) are shown in the PBS-DAPI image. (B) Quantification and comparison of cortical layer proteins stained with the marker antibodies. Intensities were averaged across three serial section per sample for statistical analysis. Cux1 (Student’s T-test p = 0.03) and SatB2 (students T-test p = 0.0001) were significantly reduced in the Pg-OMV group. Data are presented with the median and 95% CI.

Article Snippet: Sections were washed with PBS, blocked in PBS containing 0.1% tritonX- 100, and 10% donkey serum and probed with and primary antibodies to Cux1 (Protein Tech 11733-1-AP; diluted 1:300), SatB2 (Abcam ab92446; diluted 1:100), and Ctip2 (Abcam ab-18465; diluted 1:500).

Techniques: Comparison, Staining, Marker