RBFOX2 Antibodies

RNA binding protein fox-1 homolog 2 (RBFOX2)

RNA-binding protein that modulates alternative splicing events by binding to 5'-UGCAUGU-3' elements. Prevents binding of U2AF2 into the 3'-splice website.

Regulates alternative splicing of tissue-specific exons and of differentially spliced exons during erythropoiesis (By similarity). RNA-binding protein that seems to function as a coregulatory factor of ER-alpha.

Gene Information

Human
Gene Name:	RBFOX2
Uniprot:	O43251
Entrez:	23543

Family

Belongs to:
No superfamily

Alternative Names

dJ106I20.3; Fox-1 homolog B; fox-1 homologue; FOX2Fox-2; fxh; hexaribonucleotide binding protein 2; Hexaribonucleotide-binding protein 2; HRNBP2FOX-2; RBM9RTAHNRBP2; Repressor of tamoxifen transcriptional activity; RNA binding motif protein 9; RNA binding protein fox-1 homolog 2; RNA binding protein, fox-1 homolog (C. elegans) 2; RNA-binding motif protein 9; RNA-binding protein 9

Molecular Weight

Mass (kDA):
41.374 kDA

Genomic Context

Human
Location:	22q12.3
Sequence:	22; NC_000022.11 (35738734..36028892, complement)

Subcellular Localizations

Nucleus. Cytoplasm.

Diseases

• Malignant Neoplasms
• Malignant Neoplasm Of Breast
• Sclerosis
• Scleroderma, Limited
• Ataxia
• Scleroderma
• Nervous System Disorder
• Malignant Paraganglionic Neoplasm
• Renal Tubular Acidosis
• Connective Tissue Diseases
• Diffuse Scleroderma
• Neoplasms

• Nervousness
• Mammary Neoplasms
• Malignant Neoplasm Of Ovary
• Ataxia, Spinocerebellar
• Hyperplasia
• Trematode Infections

Interacting Proteins

• ATXN1
• BOLL
• C1orf94
• CAMK2B
• DAB1

• DAZAP2
• ESR1
• HNRNPF
• MAGED1
• MAPK1IP1L

• NAF1
• PSMA3
• QKI
• RBFOX1
• RBPMS2

• RHOXF2
• RNF8
• VAC14

Pathways

• Localization
• Rna Interference
• Cell Migration
• Translation
• Myofibril Assembly
• Brain Development
• Regulation Of Cell Migration
• Neurogenesis

• Pathogenesis
• Rna Splicing

PTMs

• Phosphorylation

• Cleavage

For details, visit:
bosterbio.com/bosterbio-gene-info-cards/RBFOX2

Best Uses of the RBFOX2 Marker

The RBFOX2 Marker is expressed in a variety of cell types, including neurons. This article will discuss how it is expressed in neurons and its importance in neurodegenerative diseases. You'll also discover how the RBFOX2 Marker is used in research and medicine. Read on to learn more about this intriguing gene. This article is written for those who want to further their understanding of how neurons function and how to measure RBFOX2 expression.

RBFOX2 Marker

RBFOX2 is a neuron-specific RNA repressor and regulates e18a inclusion during pre-mRNA splicing. This gene may limit CaV2.2 currents early in development in certain neuronal populations. However, the best uses of the RBFOX2 marker involve the characterization of its function. Here is a brief overview of its functions.

RBFOX2 is a key regulator of EMT. Several studies have emphasized the role of this protein in cancer cells. In a recent study, RBFOX2 increased the mesenchymal phenotype of BC cells, where NEK2 decreased mesenchymal phenotype. Using this gene, researchers were able to identify the molecular mechanisms of EMT in human cancer cells.

RBFOX1 and RBFOX2 are expressed in multiple tissues, including the heart and brain. In addition to the heart and lungs, both genes are expressed in ovary, spleen, and kidney. RBFOX3 encodes a neuron-specific nuclear protein, which can be recognized by NeuN antibodies. Both genes may be involved in alternative polyadenylation regulation, mRNA stability, and translation. Research indicates that RBFOX1 haploinsufficiency is linked to autism, intellectual disability, and epilepsy.

RBFOX2 knockout neurons also increased the expression of the TBR1 and TBR2 genes, which are markers of postmitotic cortical plate neurons. However, TBR1 and TBR2 were not significantly different between the control and RBFOX2 knockout neurons. This suggested that the expression of these genes may be affected by non-canonical Wnt pathways.

The RBFOX2 gene is located in exon 12. Its first exon, E1A, is conserved in humans. This gene participates in alternative splicing in neural development. RBFOX2 is the second largest protein in human blood. The protein is involved in several aspects of cellular metabolism and is a key player in neuronal regeneration. It is used in cell-culture experiments to determine the activity of neuronal populations.

RNA-binding proteins bind to RNA transcripts and crosslink to them when UV irradiated cells are treated with DNase. Magnetic beads are used to pull down the RNA transcripts that contain the RBFOX1 and RBFOX2 gene. RNA transcripts are then reverse-transcribed and conventional PCR analysis is performed. This procedure is repeated several times to confirm the presence of RNA-binding proteins.

RBFOX2 is expressed in many brain tissues, including the hippocampus, striatum, and cerebellum. Deletion of RBFOX1 transcript isoforms is a key cause of neurodevelopmental disorders. Because RBFOX2 has a role in neurodevelopmental disorders, identifying these variants early in their development is important for preventing or resolving these diseases.

RNA-seq data revealed that the expression of RBFOX2 was significantly reduced in NEK2-silenced MDA-MB-231 cells. Moreover, computational analysis of the datasets revealed a strong correlation between RBFOX2 and NEK2. Furthermore, hnRNPL is another RBP with high proportion of NEK2-regulated exons.

RBFOX2 expression

The RBFOX2 gene encodes a protein required for normal Purkinje cell function. Defects in this gene reduce the expression of Rbfox proteins, which can result in different neurological disorders. Deletions of this gene result in irregular Purkinje cell physiology and disrupt cerebellar development. Furthermore, the RBFOX2 gene regulates the post-transcriptional program of brain development.

A gene deletion of the Rbfox2 gene in mice results in reduced expression of the protein. Rbfox2 is necessary for the development of mature Purkinje cells. In mice with an L7-DKO, however, Rbfox2 expression was not reduced. In addition, L7-DKO mice had a decreased firing frequency and a significantly higher ISI coefficient of variation than wild-type mice.

The mice with the RBFOX2 gene mutation exhibit abnormal morphology and posture. Male homozygotes exhibit hunched posture and fail to survive to 1 mo of age. The female homozygotes exhibit a pronounced decline in body weight, with 59% fewer nuclei labeled than wild-type females at the same age. Moreover, the mice suffer from hydrocephalus.

RBFOX2 expression in neurons

RBFOX2 is a member of the RING finger transcription factor family. Its expression is restricted to neurons, heart, and muscle. The protein is expressed primarily by neurons due to the alternative splicing of exon A53, resulting in an alternative C-terminus and frame-shift. This protein regulates alternative pre-mRNA splicing by binding to the consensus sequence (U)GCAUG.

RBFOX2 is expressed in both adult and neonatal mice, and the levels of RNA polymerase II expression in these cells are similar. The expression of TBR1 in these cells is comparable between controls and RBFOX2 knockout mice. In addition, TBR1 is upregulated in neurons of postmitotic cortical plates. It is thought that Rbfox1 and RBFOX2 may contribute to Purkinje cell pacemaking.

RBFOX1 and RBFOX2 are highly expressed in neurons of various tissues, including the heart, brain, and kidney. RBFOX3 encodes a neuron-specific nuclear protein that can be recognized by NeuN antibodies. RBFOX proteins may be involved in alternative polyadenylation regulation, as well as mRNA stability and translation. RBFOX1 haploinsufficiency is associated with autism, intellectual disability, and epilepsy.

Deletion of the L7 promoter in mouse embryos did not result in altered cerebellar development. The L7 promoter is active relatively late in the development of the mouse. By two or three weeks of age, maximal genomic recombination has already occurred. Using confocal immunofluorescence, the expression of RbFOX1 and RbFOX2 was detected in Purkinje cells. RbFOX1 and RbFOX2 expression in P20 and P70 Purkinje cells resembled wild-type Purkinje cells at these ages.

The knockout and knockdown of RbFOX2 in neurons was achieved by transfection of the cDNA containing the RBFOX2 gene with two siRNA duplexes targeting human RBFOX2 (AAU GGA CGG UUA UUAU). The cells were then co-transfected with either the siRNA or a plasmid using JetPEI.

RBFOX1 and RBFOX2 are closely linked to the etiology of neurodevelopmental disorders. These proteins regulate alternative splicing of neuronal transcripts. They also target genes related to autism spectrum disorders and the vSNARE protein VAMP1. Re-expression of Vamp1 in Rbfox1 knockout mice reversed the electrophysiological abnormalities observed in the absence of Rbfox1.

To detect the presence of RBFOX2 protein in cells, we firstly lysed the neurons in RIP buffer, a protease inhibitor cocktail. Next, the lysates were incubated with an anti-RBFOX2 antibody. After that, we used Dynabeads(r) Protein G-coated Dynabeads(r) to visualize the immunoreactive protein. Finally, we visualized the results using the ChemiDoc Touch software. We used two biological replicates for each sample.

In addition, RBFOX2 KO neurons showed increased proliferation compared to normal and AS-deficient controls. This was not due to an attenuation of the canonical Wnt pathway, as the increased mRNA did not translate to more protein in the cells. The increased expression of FRZB in RBFOX2 KO neurons could indicate changes in the non-canonical Wnt pathway.