mouse anti epha4  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: EPHA1 and EPHB4 tyrosine kinase receptors regulate epithelial morphogenesis

doi: 10.1101/2024.07.15.603563

Figure Lengend Snippet: (A) RNA expression levels of all human EPHRs in Caco-2 cells were extracted from the Human Protein Atlas database and displayed in a decreasing order. (B) Western blot analysis of endogenous EPHA1, EPHA2, EPHA4 and EPHB4 in sub-confluent (70%), confluent (100%) and post-confluent (5dpc; 5 days post-confluency) Caco-2 cells cultured as monolayers. Representative images of 3 experiments are shown. (C) Confocal images displaying a representative example of EPHA1 and EPHB4 expression in 3D Caco-2 spheroid cysts. Single cells were seeded on Matrigel and grown for 6 days. Expression of apical (EZR) and tight junction (ZO-1) markers is shown, along with nuclear DAPI staining (scale bar: 15 µm).

Article Snippet: Primary antibodies used were: goat anti-EPHA1 (R&D, 1:200, #AF638), goat anti-EPHB4 (R&D, 1:200, #AF3038), mouse anti-EPHA4 (BD Biosciences, 1:1000, #610471), mouse anti-EPHA2 (Millipore-Sigma, 1:500, #05-480), mouse anti-Actin (Cell Signaling Technology, 1:1000 #3700), mouse anti-PKCι (BD, 1:500, #610175), rabbit anti-GFP (Thermo Fisher Scientific, 1:1000 #A-11122) and rabbit anti-EFNB2 (R&D, 0.2 µg/µL, #NBP1-84830).

Techniques: RNA Expression, Western Blot, Cell Culture, Expressing, Staining

Journal: PLoS ONE

Article Title: The Small GTPase RhoA Is Required for Proper Locomotor Circuit Assembly

doi: 10.1371/journal.pone.0067015

Figure Lengend Snippet: (A) Representative images of growth cones from control and RhoA cKO cortical neurons after a 30 min incubation with preclustered ephrinB3-Fc or Fc alone. Scale bar:10 µm. (B) Quantification of the growth cone collapse response of control and RhoA cKO neurons treated with ephrinB3-Fc or Fc (*p<0.001, ANOVA followed by the Tukey test). (C) Cortical neurons from RhoA cKO mice express EphA4 at levels similar to control neurons. (D) Percentage of ephrinB3-induced growth cone collapse in rat cortical neurons pretreated or not with the ROCK inhibitor Y-27632 (*p<0.001, ANOVA followed by the Tukey test). (E) Expression of ephrinB3 is lost from the midline at L2 in RhoA cKO mice. Arrowheads show ephrinB3 positive staining in the spinal cord of P5 mice. Scale bar: 10 µm. Three sections per animal and N = 3 animals were analyzed per genotype.

Article Snippet: The following purchased antibodies were used in our study: rabbit polyclonal antibodies anti-RhoA, anti-RhoB, and anti-RhoC (Cell Signaling Technology, Beverly, MA); mouse monoclonal anti-EphA4 receptor and rabbit polyclonal anti-ephrinB3 (Invitrogen, Camarillo, CA); anti-GAPDH antibody (Millipore, Temecula, CA); and the mouse monoclonal anti-acetylated tubulin antibody (Sigma, St. Louis, MO).

Techniques: Incubation, Expressing, Staining

Journal: PLoS ONE

Article Title: The Small GTPase RhoA Is Required for Proper Locomotor Circuit Assembly

doi: 10.1371/journal.pone.0067015

Figure Lengend Snippet: In control mice, when EphA4-expressing CST and spinal interneuron (IN) axons encounter ephrinB3 at the spinal cord midline, they are repulsed due to the initiation of signaling pathways that involve RhoA activation and Rac1 inhibition. Loss of RhoA causes aberrant midline crossing of CST and spinal IN axons due to a failure of neurons to retract their axons and/or the absence of ephrinB3 expression at the midline.

Article Snippet: The following purchased antibodies were used in our study: rabbit polyclonal antibodies anti-RhoA, anti-RhoB, and anti-RhoC (Cell Signaling Technology, Beverly, MA); mouse monoclonal anti-EphA4 receptor and rabbit polyclonal anti-ephrinB3 (Invitrogen, Camarillo, CA); anti-GAPDH antibody (Millipore, Temecula, CA); and the mouse monoclonal anti-acetylated tubulin antibody (Sigma, St. Louis, MO).

Techniques: Expressing, Activation Assay, Inhibition

Journal: Cell reports

Article Title: Tracheal separation is driven by NKX2-1-mediated repression of Efnb2 and regulation of endodermal cell sorting

doi: 10.1016/j.celrep.2022.110510

Figure Lengend Snippet:

Article Snippet: Immunofluorescence was performed according to standard techniques with antibodies against acetylated α-tubulin (1:250, Sigma, T7451), BrdU (1:150, Abcam, ab6326), cleaved caspase3 (1:200, Cell Signaling, 9661), EPHA4 (1:75, R&D, AF641), EPHB2 (1:75, R&D, AF467), EPHB3 (1:75, R&D, AF432), EPHB4 (1:75, R&D, AF446), phospho-EPHA2+A3+A4 (1:150, Abcam, ab62256), phospho-EPHB1+B2 (1:150, Abcam, ab61791), GFP (1:500, Abcam, ab13970), KRT5 (1:250, BioLegend, 905501), KRT8 (1:75, DSHB, AB_531826), LRIG1 (1:200, R&D, AF3688), MUC5B (1:300, Santa Cruz Biotechnology, sc-20119), NKX2-1 (1:150, Millipore, 07–601), NKX2-1 (1:100, Santa Cruz Biotechnology, sc-8761), NKX2-1 (1:300, ThermoFisher, MS-699), RFP (1:250, Abcam, ab62341), SOX2 (1:300, Neuromics, GT15098), and SOX2 (1:250, Seven Hills Bioreagents, WRAB-1236).

Techniques: Recombinant, Transfection, RNAscope, Multiplex Assay, Reporter Assay