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Sino Biological human unc5b cdna

<t>UNC5B</t> mediates FLRT2-promoted THP-1 cell maturation into macrophages. FLRT2 was overexpressed by transfecting the hFLRT2 vector, and UNC5B was silenced by transfecting UNC5B-specific shRNA into THP-1 cells. (A) Representative phase-contrast light microscopy images showing morphological alterations of THP-1 cells transfected as indicated (n = 3). Scale bar, 50 μm. (B) qPCR analysis of Itgam , Csf1r , Cd36 , Msr1 , and Olr1 mRNA in THP-1 cells transfected as indicated (n = 3). (C) Immunoblot analysis of CD36, SR-A, UNC5B, and GFP-FLRT2 proteins in THP-1 cells transfected as indicated (n = 3). (D) HUVECs expressing RFP were co-cultured with THP-1 cells transfected as indicated. After 6 h co-culture, pictures were taken using a fluorescence microscope (n = 3). Scale bar, 5 mm. (E) Transwell cell migration assay was performed in THP-1 cells transfected as indicated, and the numbers of the migrated cells were counted (n = 3). Scale bar, 100 μm. (F) Phagocytosis assays were performed by culturing the cells transfected as indicated in Texas red-conjugated zymosan particles for 2 h at 37°C (n = 3). Cells were observed for internalization of the particles by fluorescence microscopy. Scale bar, 50 μm. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Human Unc5b Cdna, supplied by Sino Biological, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+unc5b+cdna/Human+UNC5B+%2F+UNC5H2+Gene+ORF+cDNA+clone+in+cloning+vector/pmc10117657-58-0-7
Average 91 stars, based on 1 article reviews

human unc5b cdna - by Bioz Stars, 2026-10

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1) Product Images from "Fibronectin leucine-rich transmembrane protein 2 drives monocyte differentiation into macrophages via the UNC5B-Akt/mTOR axis"

Article Title: Fibronectin leucine-rich transmembrane protein 2 drives monocyte differentiation into macrophages via the UNC5B-Akt/mTOR axis

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2023.1162004

UNC5B mediates FLRT2-promoted THP-1 cell maturation into macrophages. FLRT2 was overexpressed by transfecting the hFLRT2 vector, and UNC5B was silenced by transfecting UNC5B-specific shRNA into THP-1 cells. (A) Representative phase-contrast light microscopy images showing morphological alterations of THP-1 cells transfected as indicated (n = 3). Scale bar, 50 μm. (B) qPCR analysis of Itgam , Csf1r , Cd36 , Msr1 , and Olr1 mRNA in THP-1 cells transfected as indicated (n = 3). (C) Immunoblot analysis of CD36, SR-A, UNC5B, and GFP-FLRT2 proteins in THP-1 cells transfected as indicated (n = 3). (D) HUVECs expressing RFP were co-cultured with THP-1 cells transfected as indicated. After 6 h co-culture, pictures were taken using a fluorescence microscope (n = 3). Scale bar, 5 mm. (E) Transwell cell migration assay was performed in THP-1 cells transfected as indicated, and the numbers of the migrated cells were counted (n = 3). Scale bar, 100 μm. (F) Phagocytosis assays were performed by culturing the cells transfected as indicated in Texas red-conjugated zymosan particles for 2 h at 37°C (n = 3). Cells were observed for internalization of the particles by fluorescence microscopy. Scale bar, 50 μm. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Figure Legend Snippet: UNC5B mediates FLRT2-promoted THP-1 cell maturation into macrophages. FLRT2 was overexpressed by transfecting the hFLRT2 vector, and UNC5B was silenced by transfecting UNC5B-specific shRNA into THP-1 cells. (A) Representative phase-contrast light microscopy images showing morphological alterations of THP-1 cells transfected as indicated (n = 3). Scale bar, 50 μm. (B) qPCR analysis of Itgam , Csf1r , Cd36 , Msr1 , and Olr1 mRNA in THP-1 cells transfected as indicated (n = 3). (C) Immunoblot analysis of CD36, SR-A, UNC5B, and GFP-FLRT2 proteins in THP-1 cells transfected as indicated (n = 3). (D) HUVECs expressing RFP were co-cultured with THP-1 cells transfected as indicated. After 6 h co-culture, pictures were taken using a fluorescence microscope (n = 3). Scale bar, 5 mm. (E) Transwell cell migration assay was performed in THP-1 cells transfected as indicated, and the numbers of the migrated cells were counted (n = 3). Scale bar, 100 μm. (F) Phagocytosis assays were performed by culturing the cells transfected as indicated in Texas red-conjugated zymosan particles for 2 h at 37°C (n = 3). Cells were observed for internalization of the particles by fluorescence microscopy. Scale bar, 50 μm. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Techniques Used: Plasmid Preparation, shRNA, Light Microscopy, Transfection, Western Blot, Expressing, Cell Culture, Co-Culture Assay, Fluorescence, Microscopy, Cell Migration Assay, Two Tailed Test

The direct interaction of the FLRT2 ECD with UNC5B is required for FLRT2-promoted THP-1 cell differentiation into macrophages. (A) Myc-tagged hUNC5B and/or Flag-tagged hFLRT2 were transfected into HEK293T cells. Immunoprecipitation samples were collected using anti-Flag antibody, followed by immunoblotting for Myc and Flag (n = 3). (B) Immunolabeling of FLRT2 and UNC5B in PMA-treated THP-1 cells showing the colocalization of endogenous UNC5B and FLRT2. Scale bar, 10 μm. (C) UNC5B was immunoprecipitated from PMA-incubated THP-1 cells, followed by immunoblotting for FLRT2 and UNC5B (n = 3). (D) Schematic illustration of FLRT2 and UNC5B constructs. LRRNT, leucine-rich repeat N-terminal domain. LRR, leucine-rich repeat. LRRCT, leucine-rich repeat C-terminal domain. FNIII, fibronectin type III domain. (E) UNC5B interacts with hFLRT2-ECD but not hFLRT2-ICD (n = 3). (F, G) THP-1 cells were transfected with control, human FLRT2 extracellular domain (hFLRT2-ECD) or human FLRT2 intracellular domain (hFLRT2-ICD) vector for 48 h, followed by qPCR (F) and immunoblot (G) analyses. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. NS, not significant. ** P < 0.01, **** P < 0.0001.

Figure Legend Snippet: The direct interaction of the FLRT2 ECD with UNC5B is required for FLRT2-promoted THP-1 cell differentiation into macrophages. (A) Myc-tagged hUNC5B and/or Flag-tagged hFLRT2 were transfected into HEK293T cells. Immunoprecipitation samples were collected using anti-Flag antibody, followed by immunoblotting for Myc and Flag (n = 3). (B) Immunolabeling of FLRT2 and UNC5B in PMA-treated THP-1 cells showing the colocalization of endogenous UNC5B and FLRT2. Scale bar, 10 μm. (C) UNC5B was immunoprecipitated from PMA-incubated THP-1 cells, followed by immunoblotting for FLRT2 and UNC5B (n = 3). (D) Schematic illustration of FLRT2 and UNC5B constructs. LRRNT, leucine-rich repeat N-terminal domain. LRR, leucine-rich repeat. LRRCT, leucine-rich repeat C-terminal domain. FNIII, fibronectin type III domain. (E) UNC5B interacts with hFLRT2-ECD but not hFLRT2-ICD (n = 3). (F, G) THP-1 cells were transfected with control, human FLRT2 extracellular domain (hFLRT2-ECD) or human FLRT2 intracellular domain (hFLRT2-ICD) vector for 48 h, followed by qPCR (F) and immunoblot (G) analyses. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. NS, not significant. ** P < 0.01, **** P < 0.0001.

Techniques Used: Cell Differentiation, Transfection, Immunoprecipitation, Western Blot, Immunolabeling, Incubation, Construct, Control, Plasmid Preparation, Two Tailed Test

FLRT2 activates Akt/mTOR signaling via the binding of UNC5B with Rac1. (A) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the upregulated differentially expressed genes (DEGs) in PMs of Flrt2 fl/fl and Flrt2 ΔMyel mice. (B) Immunoblot analysis of p-Akt (Ser473), p-Akt (Thr308), Akt, p-mTOR (Ser2481), mTOR, p-4E-BP-1 (Thr37/46), 4E-BP-1, p-S6K (Thr389), S6K, p-S6 (Ser240/244), and S6 proteins in PMs isolated from Flrt2 fl/fl and Flrt2 ΔMyel mice i.p. injected with thioglycollate for 3 days (n = 3 mice per group). (C) Immunoblot analysis of p-Akt (Ser473), p-Akt (Thr308), Akt, p-mTOR (Ser2481), mTOR, p-4E-BP-1 (Thr37/46), 4E-BP-1, p-S6K (Thr389), S6K, p-S6 (Ser240/244), S6, Flag-FLRT2, and FLRT2 protein levels in THP-1 cells transfected with control or hFLRT2 vector for 48 h (n = 3). (D) Immunoblot analysis of the indicated protein levels in THP-1 cells transfected with shNC or shFLRT2 #3 vector for 48 h (n = 3). (E) Immunoprecipitation samples were collected using anti-Flag antibody from control vector and Flag-UNC5B-overexpressing HEK293T cells and subjected to mass spectrometry (MS) to identify potential binding partners of UNC5B protein. A unique peptide of Rac1 protein was identified in the UNC5B-overexpressing immunoprecipitation samples by analyzing the mass-to-charge ratio of the samples. (F) The endogenous interaction between Rac1 and UNC5B was confirmed in THP-1 cells by co-immunoprecipitation (co-IP) analysis. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Figure Legend Snippet: FLRT2 activates Akt/mTOR signaling via the binding of UNC5B with Rac1. (A) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the upregulated differentially expressed genes (DEGs) in PMs of Flrt2 fl/fl and Flrt2 ΔMyel mice. (B) Immunoblot analysis of p-Akt (Ser473), p-Akt (Thr308), Akt, p-mTOR (Ser2481), mTOR, p-4E-BP-1 (Thr37/46), 4E-BP-1, p-S6K (Thr389), S6K, p-S6 (Ser240/244), and S6 proteins in PMs isolated from Flrt2 fl/fl and Flrt2 ΔMyel mice i.p. injected with thioglycollate for 3 days (n = 3 mice per group). (C) Immunoblot analysis of p-Akt (Ser473), p-Akt (Thr308), Akt, p-mTOR (Ser2481), mTOR, p-4E-BP-1 (Thr37/46), 4E-BP-1, p-S6K (Thr389), S6K, p-S6 (Ser240/244), S6, Flag-FLRT2, and FLRT2 protein levels in THP-1 cells transfected with control or hFLRT2 vector for 48 h (n = 3). (D) Immunoblot analysis of the indicated protein levels in THP-1 cells transfected with shNC or shFLRT2 #3 vector for 48 h (n = 3). (E) Immunoprecipitation samples were collected using anti-Flag antibody from control vector and Flag-UNC5B-overexpressing HEK293T cells and subjected to mass spectrometry (MS) to identify potential binding partners of UNC5B protein. A unique peptide of Rac1 protein was identified in the UNC5B-overexpressing immunoprecipitation samples by analyzing the mass-to-charge ratio of the samples. (F) The endogenous interaction between Rac1 and UNC5B was confirmed in THP-1 cells by co-immunoprecipitation (co-IP) analysis. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Techniques Used: Binding Assay, Western Blot, Isolation, Injection, Transfection, Control, Plasmid Preparation, Immunoprecipitation, Mass Spectrometry, Co-Immunoprecipitation Assay, Two Tailed Test

Schematic diagram of our major findings. Pro-differentiation factors, such as PMA, M-CSF, and thioglycollate, upregulate Flrt2 expression in monocytes. FLRT2, in turn, accelerates monocyte-to-macrophage differentiation by binding to UNC5B via its ECD and subsequently activating the Akt/mTOR signaling pathway.

Figure Legend Snippet: Schematic diagram of our major findings. Pro-differentiation factors, such as PMA, M-CSF, and thioglycollate, upregulate Flrt2 expression in monocytes. FLRT2, in turn, accelerates monocyte-to-macrophage differentiation by binding to UNC5B via its ECD and subsequently activating the Akt/mTOR signaling pathway.

Techniques Used: Expressing, Binding Assay

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Article Title: Fibronectin leucine-rich transmembrane protein 2 drives monocyte differentiation into macrophages via the UNC5B-Akt/mTOR axis
Article Snippet: .. Human UNC5B cDNA (HG13606-G) was purchased from Sino Biological and cloned into the BamHI site of the pKmyc vector. ..

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Journal: Frontiers in Immunology

Article Title: Fibronectin leucine-rich transmembrane protein 2 drives monocyte differentiation into macrophages via the UNC5B-Akt/mTOR axis

doi: 10.3389/fimmu.2023.1162004

Figure Lengend Snippet: UNC5B mediates FLRT2-promoted THP-1 cell maturation into macrophages. FLRT2 was overexpressed by transfecting the hFLRT2 vector, and UNC5B was silenced by transfecting UNC5B-specific shRNA into THP-1 cells. (A) Representative phase-contrast light microscopy images showing morphological alterations of THP-1 cells transfected as indicated (n = 3). Scale bar, 50 μm. (B) qPCR analysis of Itgam , Csf1r , Cd36 , Msr1 , and Olr1 mRNA in THP-1 cells transfected as indicated (n = 3). (C) Immunoblot analysis of CD36, SR-A, UNC5B, and GFP-FLRT2 proteins in THP-1 cells transfected as indicated (n = 3). (D) HUVECs expressing RFP were co-cultured with THP-1 cells transfected as indicated. After 6 h co-culture, pictures were taken using a fluorescence microscope (n = 3). Scale bar, 5 mm. (E) Transwell cell migration assay was performed in THP-1 cells transfected as indicated, and the numbers of the migrated cells were counted (n = 3). Scale bar, 100 μm. (F) Phagocytosis assays were performed by culturing the cells transfected as indicated in Texas red-conjugated zymosan particles for 2 h at 37°C (n = 3). Cells were observed for internalization of the particles by fluorescence microscopy. Scale bar, 50 μm. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human UNC5B cDNA (HG13606-G) was purchased from Sino Biological and cloned into the BamHI site of the pKmyc vector.

Techniques: Plasmid Preparation, shRNA, Light Microscopy, Transfection, Western Blot, Expressing, Cell Culture, Co-Culture Assay, Fluorescence, Microscopy, Cell Migration Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Fibronectin leucine-rich transmembrane protein 2 drives monocyte differentiation into macrophages via the UNC5B-Akt/mTOR axis

doi: 10.3389/fimmu.2023.1162004

Figure Lengend Snippet: The direct interaction of the FLRT2 ECD with UNC5B is required for FLRT2-promoted THP-1 cell differentiation into macrophages. (A) Myc-tagged hUNC5B and/or Flag-tagged hFLRT2 were transfected into HEK293T cells. Immunoprecipitation samples were collected using anti-Flag antibody, followed by immunoblotting for Myc and Flag (n = 3). (B) Immunolabeling of FLRT2 and UNC5B in PMA-treated THP-1 cells showing the colocalization of endogenous UNC5B and FLRT2. Scale bar, 10 μm. (C) UNC5B was immunoprecipitated from PMA-incubated THP-1 cells, followed by immunoblotting for FLRT2 and UNC5B (n = 3). (D) Schematic illustration of FLRT2 and UNC5B constructs. LRRNT, leucine-rich repeat N-terminal domain. LRR, leucine-rich repeat. LRRCT, leucine-rich repeat C-terminal domain. FNIII, fibronectin type III domain. (E) UNC5B interacts with hFLRT2-ECD but not hFLRT2-ICD (n = 3). (F, G) THP-1 cells were transfected with control, human FLRT2 extracellular domain (hFLRT2-ECD) or human FLRT2 intracellular domain (hFLRT2-ICD) vector for 48 h, followed by qPCR (F) and immunoblot (G) analyses. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. NS, not significant. ** P < 0.01, **** P < 0.0001.

Article Snippet: Human UNC5B cDNA (HG13606-G) was purchased from Sino Biological and cloned into the BamHI site of the pKmyc vector.

Techniques: Cell Differentiation, Transfection, Immunoprecipitation, Western Blot, Immunolabeling, Incubation, Construct, Control, Plasmid Preparation, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Fibronectin leucine-rich transmembrane protein 2 drives monocyte differentiation into macrophages via the UNC5B-Akt/mTOR axis

doi: 10.3389/fimmu.2023.1162004

Figure Lengend Snippet: FLRT2 activates Akt/mTOR signaling via the binding of UNC5B with Rac1. (A) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the upregulated differentially expressed genes (DEGs) in PMs of Flrt2 fl/fl and Flrt2 ΔMyel mice. (B) Immunoblot analysis of p-Akt (Ser473), p-Akt (Thr308), Akt, p-mTOR (Ser2481), mTOR, p-4E-BP-1 (Thr37/46), 4E-BP-1, p-S6K (Thr389), S6K, p-S6 (Ser240/244), and S6 proteins in PMs isolated from Flrt2 fl/fl and Flrt2 ΔMyel mice i.p. injected with thioglycollate for 3 days (n = 3 mice per group). (C) Immunoblot analysis of p-Akt (Ser473), p-Akt (Thr308), Akt, p-mTOR (Ser2481), mTOR, p-4E-BP-1 (Thr37/46), 4E-BP-1, p-S6K (Thr389), S6K, p-S6 (Ser240/244), S6, Flag-FLRT2, and FLRT2 protein levels in THP-1 cells transfected with control or hFLRT2 vector for 48 h (n = 3). (D) Immunoblot analysis of the indicated protein levels in THP-1 cells transfected with shNC or shFLRT2 #3 vector for 48 h (n = 3). (E) Immunoprecipitation samples were collected using anti-Flag antibody from control vector and Flag-UNC5B-overexpressing HEK293T cells and subjected to mass spectrometry (MS) to identify potential binding partners of UNC5B protein. A unique peptide of Rac1 protein was identified in the UNC5B-overexpressing immunoprecipitation samples by analyzing the mass-to-charge ratio of the samples. (F) The endogenous interaction between Rac1 and UNC5B was confirmed in THP-1 cells by co-immunoprecipitation (co-IP) analysis. Data are means ± SD. P values were determined using unpaired, two-tailed Student’s t -tests. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human UNC5B cDNA (HG13606-G) was purchased from Sino Biological and cloned into the BamHI site of the pKmyc vector.

Techniques: Binding Assay, Western Blot, Isolation, Injection, Transfection, Control, Plasmid Preparation, Immunoprecipitation, Mass Spectrometry, Co-Immunoprecipitation Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Fibronectin leucine-rich transmembrane protein 2 drives monocyte differentiation into macrophages via the UNC5B-Akt/mTOR axis

doi: 10.3389/fimmu.2023.1162004

Figure Lengend Snippet: Schematic diagram of our major findings. Pro-differentiation factors, such as PMA, M-CSF, and thioglycollate, upregulate Flrt2 expression in monocytes. FLRT2, in turn, accelerates monocyte-to-macrophage differentiation by binding to UNC5B via its ECD and subsequently activating the Akt/mTOR signaling pathway.

Article Snippet: Human UNC5B cDNA (HG13606-G) was purchased from Sino Biological and cloned into the BamHI site of the pKmyc vector.

Techniques: Expressing, Binding Assay

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human unc5b cdna (Sino Biological)

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