antibodie mfsd2a  (Bioss)

Journal: eLife

Article Title: Syncytin-mediated open-ended membrane tubular connections facilitate the intercellular transfer of cargos including Cas9 protein

doi: 10.7554/eLife.84391

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-MFSD2A (rabbit polyclonal) , OriGene Technologies , Cat# TA351394 , WB (1:1000) IF (1:300).

Techniques: Cell Culture, Transfection, Construct, Transduction, Recombinant, Plasmid Preparation, shRNA, Isolation, Luciferase, Reporter Assay, SYBR Green Assay, Confocal Microscopy, Software

Journal: Aging (Albany NY)

Article Title: The prognostic value of major facilitator superfamily domain-containing protein 2A in patients with hepatocellular carcinoma

doi: 10.18632/aging.102333

Figure Lengend Snippet: Decreased expression of MFSD2A in HCC. ( A ) The expression of MFSD2A in HCC and normal liver tissues was analyzed in the TCGA and GTEx databases ( P < 0.01). ( B ) RT-qPCR showed that the relative mRNA expression of MFSD2A in HCC tissues was decreased compared with that in the matched adjacent nontumorous tissues (n = 24, *P = 0.016). ( C ) Densitometric analysis of MFSD2A protein levels relative to GAPDH in HCC and corresponding normal liver samples. The expression of MFSD2A was reduced in tumor tissues when compared with that in corresponding nontumorous tissues (n = 11, *P = 0.0472). ( D ) The protein level of MFSD2A in HCC and corresponding nontumorous specimens was tested by western blotting. GAPDH was used as a loading control. RT-qPCR ( E ) and western blotting ( F ) were used to analyze the expression of MFSD2A in several HCC cell lines and one immortalized hepatic cell line LO2.

Article Snippet: After washing with PBS, the slide was incubated overnight with polyclonal goat anti-human MFSD2A antibody (Prosci Company, diluted 1/300) at 4°C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Aging (Albany NY)

Article Title: The prognostic value of major facilitator superfamily domain-containing protein 2A in patients with hepatocellular carcinoma

doi: 10.18632/aging.102333

Figure Lengend Snippet: MFSD2A expression and clinical values. ( A , B ) High expression level of MFSD2A (200×magnification); ( C ) low expression level of MFSD2A (200×magnification); ( D ) negative expression of MFSD2A (200×magnification). Low expression of MFSD2A was significantly correlated with tumor grade ( E ) and stage ( F ) analyzed with UALCAN database. The lower expression level of MFSD2A represented higher stage and pathological grade. ( G ) Patients expressing higher levels of MFSD2A show significantly better five-year overall survival ( P = 0.021). Survival curves of 79 HCC patients with different MFSD2A expression are shown. Kaplan-Meier survival curves for high expression of MFSD2A group were significantly different ( P =0.021, log-rank test) from low MFSD2A expression group in 79 HCC patients. Diagnostic outcomes for plasma MFSD2A in the diagnosis of HCC ( H ) MFSD2A concentrations in plasma. ( I ) ROC curve for MFSD2A with HCC versus HC and CHB. MFSD2A=Major Facilitator Superfamily Domain Containing 2A, HCC=hepatocellular carcinoma, CHB=chronic hepatitis B virus infection, HC=healthy control.

Article Snippet: After washing with PBS, the slide was incubated overnight with polyclonal goat anti-human MFSD2A antibody (Prosci Company, diluted 1/300) at 4°C.

Techniques: Expressing, Diagnostic Assay, Clinical Proteomics, Biomarker Discovery, Virus, Infection, Control

Journal: Aging (Albany NY)

Article Title: The prognostic value of major facilitator superfamily domain-containing protein 2A in patients with hepatocellular carcinoma

doi: 10.18632/aging.102333

Figure Lengend Snippet: Relationship between MFSD2A expression and clinicopathological features of hepatocellular carcinoma patients.

Article Snippet: After washing with PBS, the slide was incubated overnight with polyclonal goat anti-human MFSD2A antibody (Prosci Company, diluted 1/300) at 4°C.

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: The prognostic value of major facilitator superfamily domain-containing protein 2A in patients with hepatocellular carcinoma

doi: 10.18632/aging.102333

Figure Lengend Snippet: Univariate and multivariate analyses showing the overall survival rate for hepatocellular patients.

Article Snippet: After washing with PBS, the slide was incubated overnight with polyclonal goat anti-human MFSD2A antibody (Prosci Company, diluted 1/300) at 4°C.

Techniques: Expressing

Journal: Neuron

Article Title: Blood-brain barrier permeability is regulated by lipid transport-dependent suppression of caveolae-mediated transcytosis

doi: 10.1016/j.neuron.2017.03.043

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal α-Mfsd2a , Cell Signaling Technologies , Under Development; Cat#Mfsd2a14-87; RRID:AB_2617168.

Techniques: Virus, Recombinant, Sequencing, Plasmid Preparation, Software, CRISPR, Permeability

Journal: Oncotarget

Article Title: Cytotoxic stress induces transfer of mitochondria-associated human endogenous retroviral RNA and proteins between cancer cells

doi: 10.18632/oncotarget.21606

Figure Lengend Snippet: Panel A: controls. A1 : Mitochondrial Cox IV was detected in whole cell extracts, nuclear and mitochondrial fractions. A2 : lamin A & C markers were detected only in nuclear fractions indicating the absence of nuclear contaminations in the mitochondrial fraction except for cytoplasmic preparations. Panel A3 reflects the content of syncytin 2 (HERV-FRD 1 ) in all sub-cellular preparations with remarkable expression in the mitochondrial fraction of U87 RETO cells. Panel A4 : detection of the receptor for syncytin 2, MFSD2 in subcellular fractions except for cytoplasmic preparations. Panel B minor expression of pro-apoptotic proteins like BAD and BAX in U87 RETO cells. In contrast, anti-apoptotic proteins like Bcl-2 and Bcl-xL were found strongly expressed. (For comparison of cytoplasmic vs. whole cell distributions of BAX in untreated vs. treated cells, showing no difference upon etoposide-incubation, see .) Panel C : controls: isolated mitochondria with mitochondrial markers MFN1 and MFN2 (positive control) almost without extra-mitochondrial membrane proteins like ABCG2 and lamin A+C (negative controls). Panel D1 : expression of different HERV proteins in the mitochondrial fraction of U87 RETO cells. For syncytin 1 (HERV-W E1 , upper bands) the 53 kDa surface protein and an additional splicing variant below is shown. In addition, the 24 kDa band reflects the transmembrane protein of syncytin 1. For syncytin 2 the 24 kDa protein was not detectable. HERV-V 3-1 was detectable as the expected single band at 32 kDa. Panel D2 : analysis of intracellular distribution of syncytin 1 in untreated U87 RETO cells (control, left lanes) vs. treated U87 RETO cells after incubation with 5 μg/ml etoposide for 10 days. HERV proteins were found enhanced in the mitochondrial fraction upon etoposide stress. Comparable results were obtained for syncytin 2, see . Panel E : after incubating U87 RETO cells with 5 μg/ml etoposide for 10 days, receptors for syncytin 1 and 2 were also clearly detectable in mitochondrial fraction (see also panel A4 ). For SCL1A5, the endogenous protein (53 kDa) and various glycosylated forms were detected. The receptor for syncytin 2, MFSD2, was also detectable as a single band. The figure is representative of at least n = 3 independent experiments.

Article Snippet: Primary antibodies were purchased as follows: anti-HERV-WE 1 (syncytin 1) and anti-HERV-FRD 1 (syncytin 2) from Bioss, USA (Cat. No. bs2962R and bs15466R) and Biorbyt, UK (Cat. No. orb111912); anti-lamin A+C from Novus Biologicals, USA (Cat. No. EPR4100); Cox IV (Cat. No. 11967), ABCG2 (Cat. No. 4477), MFN1 (Cat. No. 13196), MFN2 (Cat. No.9482), Bcl-2 (Cat. No. 2870), Bcl-X L (Cat. No.2764), BAD (Cat. No.9239) and BAX (Cat. No.5023) primary antibodies from Cell Signaling Technology, (USA); polyclonal antibodies targeting SLC1A5 (Cat. No. bs-0473R) and MFSD2A (Cat. No. bs-6073R) from Bioss, USA; conjugated secondary antibodies from Cell Signaling (USA) and Jackson ImmunoResearch Europe Ltd. (Suffolk, UK).

Techniques: Expressing, Incubation, Isolation, Positive Control, Variant Assay

Journal: Oncotarget

Article Title: Cytotoxic stress induces transfer of mitochondria-associated human endogenous retroviral RNA and proteins between cancer cells

doi: 10.18632/oncotarget.21606

Figure Lengend Snippet: Mitochondrial samples were dual labeled for Cox IV and different HERV-proteins. Panel A depicts the expression of syncytin 1 (HERV-WE 1, A1 ), syncytin 2 (HERV-FRD 1 , A2 ), their respective receptors SLC1A5 (A3) and MFSD2 (A4) in U87 cells, which were used as source of mitochondria. The total HERV-protein expression in whole U87 cells was set to 100 %, and the relative amount of HERV-proteins associated with the mitochondrial fraction was given in relationship to the total HERV-protein amount in whole cells. Panel B depicts the measurement of different HERVs in the mitochondrial fraction of these U87 glioblastoma cells. The Cox IV- positive gated populations represent more than 94 % (picture B2 ). Picture B3 represents the negative control (secondary antibody). Both syncytin 1 and syncytin 2 ( B4 & B5 ) were found to be highly expressed, accounting for more than 98 %. HERV-V 3-1 ( B6 ) was detected in more than 67 % of the Cox IV positive populations. Plots are representative of at least n = 3 experiments.

Article Snippet: Primary antibodies were purchased as follows: anti-HERV-WE 1 (syncytin 1) and anti-HERV-FRD 1 (syncytin 2) from Bioss, USA (Cat. No. bs2962R and bs15466R) and Biorbyt, UK (Cat. No. orb111912); anti-lamin A+C from Novus Biologicals, USA (Cat. No. EPR4100); Cox IV (Cat. No. 11967), ABCG2 (Cat. No. 4477), MFN1 (Cat. No. 13196), MFN2 (Cat. No.9482), Bcl-2 (Cat. No. 2870), Bcl-X L (Cat. No.2764), BAD (Cat. No.9239) and BAX (Cat. No.5023) primary antibodies from Cell Signaling Technology, (USA); polyclonal antibodies targeting SLC1A5 (Cat. No. bs-0473R) and MFSD2A (Cat. No. bs-6073R) from Bioss, USA; conjugated secondary antibodies from Cell Signaling (USA) and Jackson ImmunoResearch Europe Ltd. (Suffolk, UK).

Techniques: Labeling, Expressing, Negative Control

Journal: Oncotarget

Article Title: Cytotoxic stress induces transfer of mitochondria-associated human endogenous retroviral RNA and proteins between cancer cells

doi: 10.18632/oncotarget.21606

Figure Lengend Snippet: Panel A 1 & 2 : positive controls, mitochondria were previously isolated from U87 RETO cells after cytotoxic stress with etoposide (as described above) and labeled with red MitoTracker ® . Endogenous mitochondria of the U87 host-cells were stained with green MitoTracker ® . After 24h-incubation without further cytotoxic stress, cellular uptake of prepared “red” mitochondria into cancer cells was clearly detectable. Panels 3-6 : experiments were performed under the same conditions as described for panels A 1 & 2 . Panels A 3 & 4 : mitochondria isolated from U87 RETO were blocked with both anti-syncytin 1 (HERV-W E1 ) and anti-syncytin 2 (HERV-FRD 1 ) blocking antibodies and the uptake of the labeled mitochondria was monitored after 24 hours. Panels A 5 & 6 : mitochondria isolated from U87 RETO were blocked with both anti-SLC1A5 and anti-MFSD2 blocking antibodies and the uptake of the labeled mitochondria was monitored after 24 hours. These results indicate that syncytin 1 and 2 as well as their receptors play a substantial role in cellular uptake of mitochondria, since the uptake of exogenous mitochondria were less perceived after blockage of these proteins. Panel B : cartoon representation of the proposed mechanisms of mitochondria entrance into U87 cells. Mitochondria from U87 RETO etoposide resistant cells were isolated and labeled with MitoTracker© red and with anti syncytin 1 or anti syncytin 2 antibodies and exogenously applied to U87 cells. The blockage of both syncytins impedes the uptake of free mitochondria.

Article Snippet: Primary antibodies were purchased as follows: anti-HERV-WE 1 (syncytin 1) and anti-HERV-FRD 1 (syncytin 2) from Bioss, USA (Cat. No. bs2962R and bs15466R) and Biorbyt, UK (Cat. No. orb111912); anti-lamin A+C from Novus Biologicals, USA (Cat. No. EPR4100); Cox IV (Cat. No. 11967), ABCG2 (Cat. No. 4477), MFN1 (Cat. No. 13196), MFN2 (Cat. No.9482), Bcl-2 (Cat. No. 2870), Bcl-X L (Cat. No.2764), BAD (Cat. No.9239) and BAX (Cat. No.5023) primary antibodies from Cell Signaling Technology, (USA); polyclonal antibodies targeting SLC1A5 (Cat. No. bs-0473R) and MFSD2A (Cat. No. bs-6073R) from Bioss, USA; conjugated secondary antibodies from Cell Signaling (USA) and Jackson ImmunoResearch Europe Ltd. (Suffolk, UK).

Techniques: Isolation, Labeling, Staining, Incubation, Blocking Assay