Anti-ENPP1 antibody [N2C2], Internal GTX103447 from GeneTex

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eLife
December 15 2020

Fig 1: Validation of select reglucosylation substrates.(A) The designated cell lines were lysed and split into whole cell lysate (WCL, 10%) or affinity purification by GST-CRT-WT or GST-CRT-Y109A and imaged by immunoblotting against the CI Man-6-Phosphate receptor. Data is representative of three independent experiments with quantification shown in panel (B). Quantifications were calculated by subtracting the value of protein in the Y109A lane from the value of protein in the associated wild-type (WT) lane, divided by the value of protein in the associated WCL lane and multiplied by 100. Error bars represent the standard deviation. Asterisks denote a p-value of less than 0.05. (C) Tandem mass tag (TMT) mass spectrometry quantification of CI Man-6-Phosphate receptor reglucosylation from ALG6/UGGT1-/- cells (Figure 2B) and ALG6/UGGT2-/- cells (Figure 2A). (D) Cartoon representation of CI Man-6-Phosphate receptor with N-glycans (branched structures), the signal sequence (gray), luminal/extracellular domain (blue), transmembrane domain (black), and intracellular domain (green) depicted. Number of amino acids and Cys residues are indicated. (E) Reglucosylation of IGF-1R, conducted as previously described above. Pro IGF-1R and mature IGF-1R are both observed due to proteolytic processing. Data are representative of three independent experiments with quantification displayed in (F). (G) TMT mass spectrometry quantification of IGF-1R from Figure 2A and B, as previously described. (H) Cartoon depiction of IGF-1R. (I) The reglucosylation of ENPP1 shown with quantification displayed in J. (K) TMT mass spectrometry quantification of ENPP1 from Figure 2A and B with cartoon depiction of ENPP1 in L. (M) Reglucosylation of ß-hexosaminidase subunit ß, conducted as previously described with quantifications displayed in N and TMT mass spectrometry quantification of ß-hexosaminidase subunit ß from Figure 2A and B in O with a cartoon depicting ß-hexosaminidase subunit ß in P.Figure 3—source data 1.Quantifications for reglucosylation validations.
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Nature communications
March 10 2017

Fig 2: Identification of membrane proteins involved in MGDAs-mediated enhancement of tumorigenic activity.(a) Summary of cDNA microarray analyses. In all, 253 genes were identified with at least twofold upregulation in MDA-MB-231, MDA-MB-361 and MDA-MB-157 compared to MDA-MB-468 cells. (b) RNAi knockdown screening of candidate genes involved in MGDAs-mediated tumorigenic activity promotion in MDA-MB-157 cells by soft agar colony formation assay. shLacZ was used as a knockdown control. (c) Expression profiles of ARMCX1, ENPP1, FRMD5, GPC6, GPR126, RFTN1 and MCT2 in six different breast cancer cell lines. The experiment was performed in technical triplicate. (d) Western blot analyses reconfirmed the expression profiles of ENPP1 and MCT2 in breast cancer cell lines (no commercial workable ARMCX1 antibody was found). The experiment in b was performed in technical triplicate and repeated at least twice with similar results. Data show means±s.d. *P<0.05 (Student's t-test).
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Nature communications
March 10 2017

Fig 3: MCT2 is required for the enhancement of tumorigenic activity associated with MGDAs.(a) Q-PCR and western blot analyses of ARMCX1, ENPP1 and MCT2 depletion and ectopic overexpression in breast cancer cells. (b) Soft agar colony formation assays showed that depletion of ARMCX1, ENPP1 and MCT2 abrogated the increase of colonies induced by MGDAs co-culture in MCF7 and MDA-MB-231 cells. (c) Soft agar colony formation assays of ARMCX1, ENPP1 and MCT2 overexpressing MDA-MB-468 and SK-BR3 cells w/ or w/o MGDAs co-culture. (d) Tumour growth assays in NOD/SCID/?null mice. MCT2-depleted MDA-MB-231 and overexpressing MDA-MB-468 cells were subcutaneously injected in both flanks of each mouse w/ or w/o MGDAs. Tumour volumes were measured every 7 days. Four mice (n=4) were used for each group. (e). Tumour weights were increased in the presence of MCT2 after co-injection with MGDAs in mouse tumorigenicity assays. The experiments in b,c were performed in technical triplicate and repeated at least twice with similar results. In b–e, data show means±s.d. *P<0.05 (Student's t-test).

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Anti-ENPP1 antibody [N2C2], Internal

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GeneTex's Anti-ENPP1 antibody [N2C2], Internal is a Rabbit Polyclonal antibody. The Anti-ENPP1 antibody [N2C2], Internal was generated using ectonucleotide pyrophosphatase/phosphodiesterase 1, ARHR2, COLED, M6S1, NPP1, NPPS, PC-1, PCA1, PDNP1 as the antigen. The Anti-ENPP1 antibody [N2C2], Internal from GeneTex is a Rabbit Polyclonal antibody to ENPP1. The Anti-ENPP1 antibody [N2C2], Internal Recognizes Human. The Anti-ENPP1 antibody [N2C2], Internal has been validated for the following Applications: WB

eLife

Nature communications

Nature communications

Anti-ENPP1 antibody [N2C2], Internal from GeneTex

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GeneTex

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Anti-ENPP1 antibody [N2C2], Internal from GeneTex

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