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Addgene inc human plxnb2 cdna

Fig. 3 <t>|</t> <t>Plexin-B2</t> affects membrane internalization in GSCs. a Dextran uptake assay of SD2 GSCs labeled with SPY-Actin. Enlarged images of boxed areas are shown below. Box plots show areas of dextran+ puncta per cell, with 25–75% quartiles, median (line), and mean (plus sign). n = 85 cells for Ctrl, n = 44 cells for PB2 KO. Mann–Whitney–Wilcoxon test, two-sided. b Top, live cell confocal plane images of SD2 GSCs with side views of z-stacks show intracellular localization of diffuse dextran- Alexa 488 in PB2 KO cells, whereas Ctrl cells contained only dextran+ endocytic vesicles. Bottom, histograms of ﬂuorescence proﬁles show bimodal distribution of dextran-Alexa 488 ﬂuorescence intensities in PB2 KO GSCs (blue and brown arrows). n = 177 cells for Ctrl, n = 161 cells for PB2 KO. Mann–Whitney–Wilcoxon test, two- sided. c, d Left, schematic of myr-palm-GFP or -CFP attached to inner membrane leaﬂet. Right, live cell images at 72 hr after transfection show ﬂuorescent probes on

Human Plxnb2 Cdna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Invasion of glioma cells through confined space requires membrane tension regulation and mechano-electrical coupling via Plexin-B2."

Article Title: Invasion of glioma cells through confined space requires membrane tension regulation and mechano-electrical coupling via Plexin-B2.

Journal: Nature communications

doi: 10.1038/s41467-024-55056-6

Fig. 3 | Plexin-B2 affects membrane internalization in GSCs. a Dextran uptake assay of SD2 GSCs labeled with SPY-Actin. Enlarged images of boxed areas are shown below. Box plots show areas of dextran+ puncta per cell, with 25–75% quartiles, median (line), and mean (plus sign). n = 85 cells for Ctrl, n = 44 cells for PB2 KO. Mann–Whitney–Wilcoxon test, two-sided. b Top, live cell confocal plane images of SD2 GSCs with side views of z-stacks show intracellular localization of diffuse dextran- Alexa 488 in PB2 KO cells, whereas Ctrl cells contained only dextran+ endocytic vesicles. Bottom, histograms of ﬂuorescence proﬁles show bimodal distribution of dextran-Alexa 488 ﬂuorescence intensities in PB2 KO GSCs (blue and brown arrows). n = 177 cells for Ctrl, n = 161 cells for PB2 KO. Mann–Whitney–Wilcoxon test, two- sided. c, d Left, schematic of myr-palm-GFP or -CFP attached to inner membrane leaﬂet. Right, live cell images at 72 hr after transfection show ﬂuorescent probes on

Figure Legend Snippet: Fig. 3 | Plexin-B2 affects membrane internalization in GSCs. a Dextran uptake assay of SD2 GSCs labeled with SPY-Actin. Enlarged images of boxed areas are shown below. Box plots show areas of dextran+ puncta per cell, with 25–75% quartiles, median (line), and mean (plus sign). n = 85 cells for Ctrl, n = 44 cells for PB2 KO. Mann–Whitney–Wilcoxon test, two-sided. b Top, live cell confocal plane images of SD2 GSCs with side views of z-stacks show intracellular localization of diffuse dextran- Alexa 488 in PB2 KO cells, whereas Ctrl cells contained only dextran+ endocytic vesicles. Bottom, histograms of ﬂuorescence proﬁles show bimodal distribution of dextran-Alexa 488 ﬂuorescence intensities in PB2 KO GSCs (blue and brown arrows). n = 177 cells for Ctrl, n = 161 cells for PB2 KO. Mann–Whitney–Wilcoxon test, two- sided. c, d Left, schematic of myr-palm-GFP or -CFP attached to inner membrane leaﬂet. Right, live cell images at 72 hr after transfection show ﬂuorescent probes on

Techniques Used: Membrane, Labeling, MANN-WHITNEY, Transfection

Fig. 6 | Plexin-B2 KO affects PIP2 localization and inner membrane surface charge in GSCs. a Left, schematic of PH(PLCD1)-GFP PIP2 probe. Right, live-cell imaging at 72 hr post transfection reveals that the PH(PLCD1)-GFP probes were largely internalized in control GSCs (arrow), but retained on membrane of PB2 KO GSCs (arrowhead). b Left, still images from videography show accumulation of PH(PLCD1)-GFP probes (arrow) in front of the nucleus (NucSpot) of migrating control SD2 in tunnels, more so in 3 than 8 µm tunnel, but not in PB2 KO cells. Dashed lines delineate cell boundary. Long arrow denotes direction of migration. Right, quantiﬁcations of the ratio of PH(PLCD1)-GFP ﬂuorescence intensity at front vs. rear of GSCs during passage. For 3 µm: n = 15 cells for WT, n = 13 cells for PB2 KO. For 8 µm: n = 13 cells for WT, n = 16 cells for PB2 KO. One-way ANOVA followed by

Figure Legend Snippet: Fig. 6 | Plexin-B2 KO affects PIP2 localization and inner membrane surface charge in GSCs. a Left, schematic of PH(PLCD1)-GFP PIP2 probe. Right, live-cell imaging at 72 hr post transfection reveals that the PH(PLCD1)-GFP probes were largely internalized in control GSCs (arrow), but retained on membrane of PB2 KO GSCs (arrowhead). b Left, still images from videography show accumulation of PH(PLCD1)-GFP probes (arrow) in front of the nucleus (NucSpot) of migrating control SD2 in tunnels, more so in 3 than 8 µm tunnel, but not in PB2 KO cells. Dashed lines delineate cell boundary. Long arrow denotes direction of migration. Right, quantiﬁcations of the ratio of PH(PLCD1)-GFP ﬂuorescence intensity at front vs. rear of GSCs during passage. For 3 µm: n = 15 cells for WT, n = 13 cells for PB2 KO. For 8 µm: n = 13 cells for WT, n = 16 cells for PB2 KO. One-way ANOVA followed by

Techniques Used: Membrane, Live Cell Imaging, Transfection, Control, Migration

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Journal: Nature communications

Article Title: Invasion of glioma cells through confined space requires membrane tension regulation and mechano-electrical coupling via Plexin-B2.

doi: 10.1038/s41467-024-55056-6

Figure Lengend Snippet: Fig. 3 | Plexin-B2 affects membrane internalization in GSCs. a Dextran uptake assay of SD2 GSCs labeled with SPY-Actin. Enlarged images of boxed areas are shown below. Box plots show areas of dextran+ puncta per cell, with 25–75% quartiles, median (line), and mean (plus sign). n = 85 cells for Ctrl, n = 44 cells for PB2 KO. Mann–Whitney–Wilcoxon test, two-sided. b Top, live cell confocal plane images of SD2 GSCs with side views of z-stacks show intracellular localization of diffuse dextran- Alexa 488 in PB2 KO cells, whereas Ctrl cells contained only dextran+ endocytic vesicles. Bottom, histograms of ﬂuorescence proﬁles show bimodal distribution of dextran-Alexa 488 ﬂuorescence intensities in PB2 KO GSCs (blue and brown arrows). n = 177 cells for Ctrl, n = 161 cells for PB2 KO. Mann–Whitney–Wilcoxon test, two- sided. c, d Left, schematic of myr-palm-GFP or -CFP attached to inner membrane leaﬂet. Right, live cell images at 72 hr after transfection show ﬂuorescent probes on

Article Snippet: The lentiviral vector for Dox-inducible Plexin-B2 overexpression was generated by inserting human PLXNB2 cDNA into a Dox controlled expression vector (pLenti-CMVtight-PLXNB2 iOE; deposited as Nature Communications | (2025) 16:272 15 Addgene #176849)19.

Techniques: Membrane, Labeling, MANN-WHITNEY, Transfection

Journal: Nature communications

Article Title: Invasion of glioma cells through confined space requires membrane tension regulation and mechano-electrical coupling via Plexin-B2.

doi: 10.1038/s41467-024-55056-6

Figure Lengend Snippet: Fig. 6 | Plexin-B2 KO affects PIP2 localization and inner membrane surface charge in GSCs. a Left, schematic of PH(PLCD1)-GFP PIP2 probe. Right, live-cell imaging at 72 hr post transfection reveals that the PH(PLCD1)-GFP probes were largely internalized in control GSCs (arrow), but retained on membrane of PB2 KO GSCs (arrowhead). b Left, still images from videography show accumulation of PH(PLCD1)-GFP probes (arrow) in front of the nucleus (NucSpot) of migrating control SD2 in tunnels, more so in 3 than 8 µm tunnel, but not in PB2 KO cells. Dashed lines delineate cell boundary. Long arrow denotes direction of migration. Right, quantiﬁcations of the ratio of PH(PLCD1)-GFP ﬂuorescence intensity at front vs. rear of GSCs during passage. For 3 µm: n = 15 cells for WT, n = 13 cells for PB2 KO. For 8 µm: n = 13 cells for WT, n = 16 cells for PB2 KO. One-way ANOVA followed by

Techniques: Membrane, Live Cell Imaging, Transfection, Control, Migration

Co-immunoprecipitation (IP) experiments showing homophilic interaction of human and murine B3 and no interaction of B3 with plexins A1, B1 or B2 . Total lysates and IPs were analyzed by Western blot (WB) using antibodies as indicated in the figures. IP was performed with pAbB3-B against human B3 and shows interaction between mouse and human B3 and no interaction between B3 and human plexins A1, B1 and B2. (A) Cells were co-transfected with pSecTag2B/B3 encoding myc-tagged human full-length B3 and pcDNA3.1/mB3 encoding V5-tagged mouse B3 lacking most of its intracellular part. Cells transfected with pcDNA3.1/mB3 and pSecTag2B vector without insert served as negative control. (B) COS-7 cells were co-transfected with pIRES/B3 encoding non-tagged full-length human B3 and pcDNAVSV/A1 encoding VSV-tagged full-length human plexin A1. Cells co-transfected with pcDNAVSV/A1 and pIRES vector without insert served as negative control. (C) COS-7 cells were co-transfected with pIRES/B3 and pcDNAVSV/B1 encoding VSV-tagged full-length human plexin B1. Cells co-transfected with pcDNAVSV/B1 and pIRES vector without insert served as negative control. (D) COS-7 cells were co-transfected with pIRES/B3 and pFLAG/B2 encoding FLAG-tagged full-length human plexin B2. Cells co-transfected with pFLAG/B2 and pIRES vector without insert served as negative control.

Journal: BMC Neuroscience

Article Title: Plexin B3 promotes neurite outgrowth, interacts homophilically, and interacts with Rin

doi: 10.1186/1471-2202-6-53

Figure Lengend Snippet: Co-immunoprecipitation (IP) experiments showing homophilic interaction of human and murine B3 and no interaction of B3 with plexins A1, B1 or B2 . Total lysates and IPs were analyzed by Western blot (WB) using antibodies as indicated in the figures. IP was performed with pAbB3-B against human B3 and shows interaction between mouse and human B3 and no interaction between B3 and human plexins A1, B1 and B2. (A) Cells were co-transfected with pSecTag2B/B3 encoding myc-tagged human full-length B3 and pcDNA3.1/mB3 encoding V5-tagged mouse B3 lacking most of its intracellular part. Cells transfected with pcDNA3.1/mB3 and pSecTag2B vector without insert served as negative control. (B) COS-7 cells were co-transfected with pIRES/B3 encoding non-tagged full-length human B3 and pcDNAVSV/A1 encoding VSV-tagged full-length human plexin A1. Cells co-transfected with pcDNAVSV/A1 and pIRES vector without insert served as negative control. (C) COS-7 cells were co-transfected with pIRES/B3 and pcDNAVSV/B1 encoding VSV-tagged full-length human plexin B1. Cells co-transfected with pcDNAVSV/B1 and pIRES vector without insert served as negative control. (D) COS-7 cells were co-transfected with pIRES/B3 and pFLAG/B2 encoding FLAG-tagged full-length human plexin B2. Cells co-transfected with pFLAG/B2 and pIRES vector without insert served as negative control.

Article Snippet: Human PLXNB2 cDNA (KIAA 0315) was kindly provided by Dr. T. Nagase (Kazusa DNA Research Institute, Kisarazu, Japan).

Techniques: Immunoprecipitation, Western Blot, Transfection, Plasmid Preparation, Negative Control

Expression of plexin B2, plexin B3 and L1 protein in stably transfected NIH-3T3 cells . Western blot detection of human L1, plexin B2 and plexin B3 protein in stably transfected NIH-3T3 substrate cells. In all cases same protein amounts were applied and non-transfected NIH-3T3 cells served as negative control. (A) Western blot detection of human L1 protein in stably pIRES/L1-tranfected NIH-3T3 cells using antibody pAbex2. (B) Western blot detection of human plexin B2 (B2) protein in stably pFLAG/B2-tranfected NIH-3T3 cells using anti-FLAG antibody. (C) Western blot detection of human plexin B3 (B3) protein in stably pIRES/B3-tranfected NIH-3T3 cells using antibody pAbB3-B.

Journal: BMC Neuroscience

Article Title: Plexin B3 promotes neurite outgrowth, interacts homophilically, and interacts with Rin

doi: 10.1186/1471-2202-6-53

Figure Lengend Snippet: Expression of plexin B2, plexin B3 and L1 protein in stably transfected NIH-3T3 cells . Western blot detection of human L1, plexin B2 and plexin B3 protein in stably transfected NIH-3T3 substrate cells. In all cases same protein amounts were applied and non-transfected NIH-3T3 cells served as negative control. (A) Western blot detection of human L1 protein in stably pIRES/L1-tranfected NIH-3T3 cells using antibody pAbex2. (B) Western blot detection of human plexin B2 (B2) protein in stably pFLAG/B2-tranfected NIH-3T3 cells using anti-FLAG antibody. (C) Western blot detection of human plexin B3 (B3) protein in stably pIRES/B3-tranfected NIH-3T3 cells using antibody pAbB3-B.

Article Snippet: Human PLXNB2 cDNA (KIAA 0315) was kindly provided by Dr. T. Nagase (Kazusa DNA Research Institute, Kisarazu, Japan).

Techniques: Expressing, Stable Transfection, Transfection, Western Blot, Negative Control

Plexins B2 and B3 stimulate neurite outgrowth . (A) Examples of primary murine cerebellar neurons isolated from six days old c57BL/6J mice and grown on L1-, B2-, or B3- expressing transfected (L1, B2, B3) or non-transfected (3T3) NIH-3T3 substrate cells. (B) Mean length of neurites of murine cerebellar neurons grown for 24 h on NIH-3T3 substrate cells. Nontransfected substrate cells (3T3) served as negative control. Cells transfected with pIRES/L1 encoding neuronal cell surface molecule L1 (L1) served as positive control. The strongest stimulation of neurite outgrowth was observed on substrate cells transfected with pIRES/B3 encoding full-length plexin B3 (B3). Plexin B2-expressing substrate cells (B2) were transfected with expression construct pFLAG/B2. Each outgrowth assay was done in three independent experiments. Pooled data of the triple experiments are shown. A minimum of 400 neurons were analyzed for each substrate cell type. Error bars: S.E. of mean × 1. Mean neurite length differed significantly between all groups (ANOVA; **, p < 0.0005; *, p = 0.0033). (C) Culmulative size distribution patterns of the neurites given in B.

Journal: BMC Neuroscience

Article Title: Plexin B3 promotes neurite outgrowth, interacts homophilically, and interacts with Rin

doi: 10.1186/1471-2202-6-53

Figure Lengend Snippet: Plexins B2 and B3 stimulate neurite outgrowth . (A) Examples of primary murine cerebellar neurons isolated from six days old c57BL/6J mice and grown on L1-, B2-, or B3- expressing transfected (L1, B2, B3) or non-transfected (3T3) NIH-3T3 substrate cells. (B) Mean length of neurites of murine cerebellar neurons grown for 24 h on NIH-3T3 substrate cells. Nontransfected substrate cells (3T3) served as negative control. Cells transfected with pIRES/L1 encoding neuronal cell surface molecule L1 (L1) served as positive control. The strongest stimulation of neurite outgrowth was observed on substrate cells transfected with pIRES/B3 encoding full-length plexin B3 (B3). Plexin B2-expressing substrate cells (B2) were transfected with expression construct pFLAG/B2. Each outgrowth assay was done in three independent experiments. Pooled data of the triple experiments are shown. A minimum of 400 neurons were analyzed for each substrate cell type. Error bars: S.E. of mean × 1. Mean neurite length differed significantly between all groups (ANOVA; **, p < 0.0005; *, p = 0.0033). (C) Culmulative size distribution patterns of the neurites given in B.

Article Snippet: Human PLXNB2 cDNA (KIAA 0315) was kindly provided by Dr. T. Nagase (Kazusa DNA Research Institute, Kisarazu, Japan).

Techniques: Isolation, Expressing, Transfection, Negative Control, Positive Control, Construct

Co-immunoprecipitation (IP) experiments showing interaction of B3 with Rin . Total lysates and IPs were analyzed by Western blot (WB) using antibodies as indicated in the figures. (A) COS-7 cells were co-transfected with pIRES/B3 encoding non-tagged full-length human B3 and pFLAG/Rin encoding FLAG-tagged full-length human Rin. Cells co-transfected with pIRES/B3 and pFLAG vector without insert served as negative control. IP was performed using anti-FLAG antibodies. (B) COS-7 cells were co-transfected with pFLAG/B2 encoding FLAG-tagged full-length human plexin B2 and pFLAG/Rin encoding FLAG-tagged full-length human Rin. Cells co-transfected with pFLAG/B2 and pFLAG vector without insert served as negative control. IP was performed using anti-Rin antibodies. Bands marked with * represent antibodies precipitated by protein-A-agarose.

Journal: BMC Neuroscience

Article Title: Plexin B3 promotes neurite outgrowth, interacts homophilically, and interacts with Rin

doi: 10.1186/1471-2202-6-53

Figure Lengend Snippet: Co-immunoprecipitation (IP) experiments showing interaction of B3 with Rin . Total lysates and IPs were analyzed by Western blot (WB) using antibodies as indicated in the figures. (A) COS-7 cells were co-transfected with pIRES/B3 encoding non-tagged full-length human B3 and pFLAG/Rin encoding FLAG-tagged full-length human Rin. Cells co-transfected with pIRES/B3 and pFLAG vector without insert served as negative control. IP was performed using anti-FLAG antibodies. (B) COS-7 cells were co-transfected with pFLAG/B2 encoding FLAG-tagged full-length human plexin B2 and pFLAG/Rin encoding FLAG-tagged full-length human Rin. Cells co-transfected with pFLAG/B2 and pFLAG vector without insert served as negative control. IP was performed using anti-Rin antibodies. Bands marked with * represent antibodies precipitated by protein-A-agarose.

Article Snippet: Human PLXNB2 cDNA (KIAA 0315) was kindly provided by Dr. T. Nagase (Kazusa DNA Research Institute, Kisarazu, Japan).

Techniques: Immunoprecipitation, Western Blot, Transfection, Plasmid Preparation, Negative Control

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