Journal: Bone research

Article Title: IRF1-mediated upregulation of PARP12 promotes cartilage degradation by inhibiting PINK1/Parkin dependent mitophagy through ISG15 attenuating ubiquitylation and SUMOylation of MFN1/2.

doi: 10.1038/s41413-024-00363-3

Figure Lengend Snippet: Fig. 5 ISG15 upregulated by PAPR12, increasing ISGylation of MFN1/2 and then attenuating the ubiquitylation and SUMOylation to inhibit PINK1/Parkin-dependent mitophagy. a Immunoblotting (IB) analysis of ubiquitylation and ISG15 in PHCs following transfection with KD-ISG15 or KD-NC. b Protein quantification of (a) using ImageJ. n = 3 per group. c IB analysis of ubiquitylation of mitochondrial proteins. Mitochondria were isolated from KD-NC and KD-ISG15 PHCs transfected with a HA-ubiquitin construct. d Protein quantification of (d) using ImageJ. n = 3 per group. e IB analysis of SUMO2/3 and ubiquitin with SUMO2/3-conjugated proteins in KD-NC and KD-ISG15 PHCs immunoprecipitated using SUMO2/3 affinity beads. f IB analysis of MFN1/2, MFN1, MFN2, ubiquitin, SUMO2/3 and ISG15 in KD-NC and KD-ISG15 PHCs treated with or without MG-132 after immunoprecipitation of MFN1/2. The molecular marker of IB plots of ubiquitin, SUMO2/3, and ISG15 conjugates with MFN1/2 is 70 kD and above when considering the molecular weight of MFN1/2 at 85 kD. g Quantification of MFN1/2, ubiquitin, SUMO2/3, and ISG15 of (f) using ImageJ after immunoprecipitation of MFN1/2. n = 3 per group. h IB analysis of MFN1/2, MFN1, MFN2, ubiquitin, SUMO2/3, and ISG15 in PHCs following either knockdown or overexpression of PARP12 after immunoprecipitation of MFN1/2. i Quantification of MFN1/2, ubiquitin, SUMO2/3, and ISG15 of (h) using ImageJ. n = 3 per group. j MitoTracker Red and LysoTracker Green staining of PHCs was observed by confocal microscopy. Scale bars: 10 µm. k Western blot analysis of PARP12, ISG15, LC3B, p62, MFN1, MFN2, PINK1, Parkin, COL2A1, aggrecan, MMP13, RUNX2, Bcl2, Bax, and NLRP3 inflammasome activity in PHCs coinfected with PARP12 KD-03 shRNA and ISG15-OE adenovirus. n = 3 per group. l ROS staining with PARP12 knockdown and ISG15 overexpression. Scale bars: 100 µm. m JC-1 staining in PHCs following transfection with PARP12 knockdown and ISG15 overexpression. Scale bars: 100 µm. Data are presented as the mean ± SD. Paired t-test (b, d, i) and one-way analysis of variance followed by Tukey’s multiple comparison test (g) were used for statistical analysis. *P < 0.05, **P < 0.01, and ***P < 0.001

Article Snippet: According to the manufacturer’s instructions of SUMOylation 2/3 affinity beads (BK162, Cytoskeleton, Inc.), SUMO2/3-conjugated proteins in chondrocytes lysates were immunoprecipitated.

Techniques: Western Blot, Transfection, Isolation, Ubiquitin Proteomics, Construct, Immunoprecipitation, Marker, Molecular Weight, Knockdown, Over Expression, Staining, Confocal Microscopy, Activity Assay, shRNA, Comparison

Journal: Bone research

Article Title: IRF1-mediated upregulation of PARP12 promotes cartilage degradation by inhibiting PINK1/Parkin dependent mitophagy through ISG15 attenuating ubiquitylation and SUMOylation of MFN1/2.

doi: 10.1038/s41413-024-00363-3

Figure Lengend Snippet: Fig. 8 PARP12 modulates osteoarthritis (OA) pathogenesis in monosodium iodoacetate (MIA)-treated rats. a Experimental diagram of the MIA OA rat model treated with XAV-939 or PARP12 overexpression (OE) adenovirus. Rats were evaluated at age of 10 weeks. n = 6 per group. b 3D reconstruction images of micro-CT scanning of the knees of rats treated with XAV-939 or PARP12-OE adenovirus. n = 5 per group. c–f Analysis of BV/TV, BS/TV, trabecular thickness, and trabecular numbers. n = 5 per group. g Western blot analysis of PARP12, COL2A1, aggrecan, MMP13, RUNX2, Bcl2/Bax, LC3B, p62, MFN1, MFN2, PINK1, Parkin and NLRP3 inflammasome activity in chondrocytes of rats treated with XAV-939 or PARP12 overexpression (OE) adenovirus. n = 3 per group. h Representative images of Safranin O and IHC staining of PARP12, COL2A1 and MMP13. Scale bars: 250 µm (first row) and 50 µm (second row). i Quantification of macroscopic score based on staining results in (h). n = 3 per group. j–l Quantification of PARP12, COL2A1, and MMP13 positive chondrocytes based on staining results in (h). n = 3 per group. m–o ROS staining, ATP level and JC-1 staining in chondrocytes of rats treated with XAV-939 or PARP12 OE adenovirus. p Schematic representation of the mechanism by which IRF1-mediated upregulation of PARP12 promotes cartilage degradation by inhibiting PINK1/Parkin dependent mitophagy through ISG15 attenuating the ubiquitylation and SUMOylation of MFN1/2. Data are presented as the mean ± SD. Paired t-test (c–f, j–l, n) and non-parametric Mann-Whitney U test (i) were used for statistical analysis. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: According to the manufacturer’s instructions of SUMOylation 2/3 affinity beads (BK162, Cytoskeleton, Inc.), SUMO2/3-conjugated proteins in chondrocytes lysates were immunoprecipitated.

Techniques: Over Expression, Micro-CT, Western Blot, Activity Assay, Immunohistochemistry, Staining, MANN-WHITNEY

Journal: Cell Communication and Signaling : CCS

Article Title: Src family kinases interfere with dimerization of STAT5A through a phosphotyrosine-SH2 domain interaction

doi: 10.1186/s12964-014-0081-7

Figure Lengend Snippet: The SH2 domain of STAT5A is required for efficient binding to Src kinases. (A) Domain structure of fluorescently labeled STAT5A-eYFP. (B) Subcellular localization of STAT5A-eYFP in the absence or presence of Epo. HeLa T-REx HA-EpoR cells stably transfected with STAT5A-eYFP were stimulated with 1 U/ml Epo for 30 min and the localization of STAT5A-eYFP was analyzed by confocal microscopy. Scale bars: 20 μm. (C) Subcellular localization of STAT5A-eYFP (upper panel), STAT5A R618Q -eYFP (middle panel) and STAT3-eYFP (lower panel) was investigated in the presence of vSrc-dsRed. HeLa T-REx vSrc-dsRed cells were treated with 5 ng/ml doxycycline and transfected with the indicated constructs and the distribution of fluorescently labeled fusion proteins was analyzed after 24 h by confocal microscopy. Scale bars: 20 μm. (D) Quantification of the relative subcellular distribution of eYFP-labeled STAT3 and STAT5A constructs in HeLa T-REx HA-EpoR cells stably expressing STAT5A-eYFP (B) and HeLa T-REx vSrc-dsRed cells transfected with STAT5A-eYFP, STAT5A R618Q -eYFP or STAT3-eYFP (C) . The expression of the HA-EpoR and vSrc-dsRed was induced with 5 ng/ml doxycycline for 24 hours. Mean fluorescence intensities (MFI) of the cytoplasm and nucleus were determined using the Zen 2012 software and changes in the ratio between the compartments were plotted. The data shown are means ± SD of n = 30 cells and were statistically evaluated by Student’s t -test. ***p < 0.0005. n.s. = not significant. (E + F) HeLa T-REx FRT cells were co-transfected with plasmids coding for STAT5A-eYFP or STAT5A R618Q -eYFP and vSrc-dsRed or Hck-dsRed. Fluorescently labeled STAT5 was immunoprecipitated from cell lysates using a GFP antibody and analyzed by immunoblotting for the presence of vSrc-dsRed or Hck-dsRed 24 h after transfection. The expression and phosphorylation of STAT5A and vSrc/Hck proteins was analyzed in the whole cellular lysates (WCL) using antibodies against pY 416 -Src, Src, Hck, pY 694/699 -STAT5A/B and GFP.

Article Snippet: Anti-pY 694/699 -STAT5A/B (#9351), anti-pY 416 -Src (#2101), anti-pY 412 -Abl (#2865), anti-Hsp70 (#4872, Cell Signaling, Beverly, USA), anti-STAT5A (clone #5073, rabbit polyclonal antiserum was kindly provided by Richard Moriggl, Ludwig Boltzmann Institute for Cancer Research (LBI-CR), Vienna, Austria), anti-GFP (600-103-215, Rockland, Gilbertsville, USA), anti-FLAG (F3165), anti-α-tubulin (T5168, Sigma, St. Louis, USA), anti-GAPDH (sc-32233), anti-Abl (sc-131), anti-Hck (sc-72), anti-cSrc (sc-19, Santa Cruz Biotechnology, Santa Cruz, USA), anti-vSrc (MABS193, Millipore, Billerica, MA, USA) and anti-HA (MMS-101R, Covance, Princeton, New Jersey, USA) antibodies were used for immunoblotting.

Techniques: Binding Assay, Labeling, Stable Transfection, Transfection, Confocal Microscopy, Construct, Expressing, Fluorescence, Software, Immunoprecipitation, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Src family kinases interfere with dimerization of STAT5A through a phosphotyrosine-SH2 domain interaction

doi: 10.1186/s12964-014-0081-7

Figure Lengend Snippet: STAT5A binds to the phosphorylated activation loop of SFK. (A) Domain structure of vSrc-dsRed. Selected amino acids are highlighted. A multiple sequence alignment of the activation loop of SFK is shown. Autophosphorylation site is highlighted (red). (*) conserved amino acids, (:) similar properties . Bold characters highlight peptide sequence used for precipitation. (B + C) HeLa T-REx FRT cells stably expressing STAT5A-eYFP were transfected with the indicated vSrc-dsRed variants. Phosphorylation was analyzed 24 h after transfection using antibodies against pY 416 -Src, Src, pY 694/699 -STAT5A/B and STAT5A. CTRL = untransfected cells. (D) Quantification of relative subcellular distribution of STAT5A in HeLa T-REx FRT stably expressing STAT5A-eYFP and the indicated vSrc-dsRed mutants. Mean fluorescence intensity (MFI) of eYFP-fluorescence in the cytoplasm and nucleus were determined using the Zen 2012 software and changes in the ratio between the compartments were plotted. Data show means ± SD of n = 10 cells and were statistically evaluated by Student’s t -test. ***p < 0.0005, **p < 0.005, *p < 0.05, n.s. = not significant. (E) HeLa T-REx FRT cells stably expressing STAT5A-eYFP were transfected with vSrc K295N -dsRed or vSrc Y416F -dsRed. Subcellular distribution of STAT5A-eYFP was analyzed 24 h after transfection by confocal microscopy. Scale bars: 20 μm. (F + G) HeLa T-REx FRT cells were co-transfected with plasmids coding for vSrc-dsRed (Hck-dsRed), vSrc K295N -dsRed (Hck K269N -dsRed) or vSrc Y416F -dsRed (Hck Y390F -dsRed) and STAT5A-eYFP. STAT5-eYFP was immunoprecipitated from cell lysates using a GFP antibody and analyzed by immunoblotting for the presence of vSrc-dsRed (Hck-dsRed) 24 h after transfection. Expression and phosphorylation of STAT5A and vSrc proteins was analyzed in the WCL using antibodies against pY 416 -Src, Src, Hck, pY 694/699 -STAT5A/B and GFP. (s) short exposure, (l) long exposure. (H) HeLa T-REx FRT cells expressing STAT5A-eYFP or STAT5A R618Q -eYFP were lysed and incubated with a Src-peptide containing tyrosine- or phosphotyrosine 416. Precipitates and WCL were analyzed by immunoblotting using a GFP-specific antibody.

Article Snippet: Anti-pY 694/699 -STAT5A/B (#9351), anti-pY 416 -Src (#2101), anti-pY 412 -Abl (#2865), anti-Hsp70 (#4872, Cell Signaling, Beverly, USA), anti-STAT5A (clone #5073, rabbit polyclonal antiserum was kindly provided by Richard Moriggl, Ludwig Boltzmann Institute for Cancer Research (LBI-CR), Vienna, Austria), anti-GFP (600-103-215, Rockland, Gilbertsville, USA), anti-FLAG (F3165), anti-α-tubulin (T5168, Sigma, St. Louis, USA), anti-GAPDH (sc-32233), anti-Abl (sc-131), anti-Hck (sc-72), anti-cSrc (sc-19, Santa Cruz Biotechnology, Santa Cruz, USA), anti-vSrc (MABS193, Millipore, Billerica, MA, USA) and anti-HA (MMS-101R, Covance, Princeton, New Jersey, USA) antibodies were used for immunoblotting.

Techniques: Activation Assay, Sequencing, Stable Transfection, Expressing, Transfection, Fluorescence, Software, Confocal Microscopy, Immunoprecipitation, Western Blot, Incubation

Journal: Cell Communication and Signaling : CCS

Article Title: Src family kinases interfere with dimerization of STAT5A through a phosphotyrosine-SH2 domain interaction

doi: 10.1186/s12964-014-0081-7

Figure Lengend Snippet: SFK-mediated cytoplasmic localization of STAT5A is dominant over BCR-ABL induced nuclear accumulation. (A) HeLa T-REx BCR-ABL cells were transiently transfected with STAT5A-eYFP and either treated with 5 ng/ml doxycycline for 24 h to induce BCR-ABL expression (lower panel) or left untreated (upper panel). Fixation was performed with methanol. Fixed cells were stained for BCR-ABL using a cABL-specific primary antibody and a secondary antibody conjugated to Alexa Fluor-405. The subcellular distribution of STAT5A-eYFP was analyzed by confocal microscopy. Scale bars: 20 μm. (B) The subcellular distribution of STAT5A-eYFP was investigated in the presence of vSrc-dsRed (upper panel), vSrc K295N -dsRed (middle panel) or vSrc Y416F -dsRed (lower panel) in HeLa T-REx BCR-ABL cells that were treated with 5 ng/ml doxycycline for 24 h. Fixation was performed with methanol. Fixed cells were stained for BCR-ABL using an Abl-specific primary antibody and a secondary antibody conjugated to Alexa Fluor-405. Scale bars: 20 μm. (C) HeLa T-REx BCR-ABL cells were co-transfected with vSrc-dsRed, or the respective kinase activity affecting mutants vSrc K295N -dsRed or vSrc Y416F -dsRed and STAT5A-eYFP. The cells were either treated with 5 ng/ml doxycycline for 24 h (lanes 1–3) to induce the expression of BCR-ABL or left untreated (lane 4). Protein expression and phosphorylation in the cellular extracts was investigated by immunoblotting with antibodies against pY 412 -cABL, cABL, pY 694/699 -STAT5A/B, STAT5A, pY 416 -Src and Src. α-Tubulin served as a loading control.

Article Snippet: Anti-pY 694/699 -STAT5A/B (#9351), anti-pY 416 -Src (#2101), anti-pY 412 -Abl (#2865), anti-Hsp70 (#4872, Cell Signaling, Beverly, USA), anti-STAT5A (clone #5073, rabbit polyclonal antiserum was kindly provided by Richard Moriggl, Ludwig Boltzmann Institute for Cancer Research (LBI-CR), Vienna, Austria), anti-GFP (600-103-215, Rockland, Gilbertsville, USA), anti-FLAG (F3165), anti-α-tubulin (T5168, Sigma, St. Louis, USA), anti-GAPDH (sc-32233), anti-Abl (sc-131), anti-Hck (sc-72), anti-cSrc (sc-19, Santa Cruz Biotechnology, Santa Cruz, USA), anti-vSrc (MABS193, Millipore, Billerica, MA, USA) and anti-HA (MMS-101R, Covance, Princeton, New Jersey, USA) antibodies were used for immunoblotting.

Techniques: Transfection, Expressing, Staining, Confocal Microscopy, Activity Assay, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Src family kinases interfere with dimerization of STAT5A through a phosphotyrosine-SH2 domain interaction

doi: 10.1186/s12964-014-0081-7

Figure Lengend Snippet: Binding of STAT5A to Src kinases interferes with dimerization. (A) HeLa T-REx HA-EpoR cells stably expressing STAT5A-eYFP were transfected with STAT5A-FLAG. The cells were treated with 5 ng/ml doxyxcyline for 24 h to induce the expression of the HA-tagged EpoR and stimulated with 5 U/ml Epo for 30 minutes or left untreated. HeLa T-REx vSrc-dsRed cells stably expressing STAT5A-eYFP were transfected with STAT5A-FLAG. The expression of vSrc-dsRed was induced for 8 h with 5 ng/ml doxycycline or the cells were left untreated. STAT5A-eYFP was immunoprecipitated from cell lysates using a GFP antibody and analyzed by immunoblotting for the presence of STAT5A-FLAG. The expression and phosphorylation of STAT5A-eYFP and STAT5A-FLAG was analyzed in the WCL using antibodies against pY 694/699 -STAT5A/B, GFP and the FLAG-tag. (B) HeLa T-REx HA-EpoR cells stably expressing STAT5A-eYFP were treated with 5 ng/ml doxyxcyline for 24 h to induce the expression of the human EpoR and stimulated with 5 U/ml Epo for 30 minutes or left untreated. HeLa T-REx vSrc-dsRed cells stably expressing STAT5A-eYFP or a STAT5A S710F -eYFP were treated with 5 ng/ml doxyxcline for 8 h or the cells were left untreated. Cellular extracts were prepared under native conditions and STAT5A-eYFP dimers were separated from monomers by blue native PAGE electrophoresis (NP). STAT5A-eYFP dimer complexes were measured by the detection of the eYFP fluorescence. The cellular extracts were subjected to immunoblotting using antibodies against pY 694/699 -STAT5A/B, STAT5A, Src and the HA-tag of the EpoR. (C) Confocal microscopy analysis of HeLa T-REx FRT cells co-expressing vSrc-dsRed together with STAT5A S710F -eYFP (upper panel), a serine phosphorylation mimicking mutant STAT5A S710D -eYFP (middle panel) or a serine phosphorylation deficient mutant STAT5A S710A -eYFP (lower panel). Methanol fixation was performed 24 h after transfection. Scale bars: 20 μm.

Article Snippet: Anti-pY 694/699 -STAT5A/B (#9351), anti-pY 416 -Src (#2101), anti-pY 412 -Abl (#2865), anti-Hsp70 (#4872, Cell Signaling, Beverly, USA), anti-STAT5A (clone #5073, rabbit polyclonal antiserum was kindly provided by Richard Moriggl, Ludwig Boltzmann Institute for Cancer Research (LBI-CR), Vienna, Austria), anti-GFP (600-103-215, Rockland, Gilbertsville, USA), anti-FLAG (F3165), anti-α-tubulin (T5168, Sigma, St. Louis, USA), anti-GAPDH (sc-32233), anti-Abl (sc-131), anti-Hck (sc-72), anti-cSrc (sc-19, Santa Cruz Biotechnology, Santa Cruz, USA), anti-vSrc (MABS193, Millipore, Billerica, MA, USA) and anti-HA (MMS-101R, Covance, Princeton, New Jersey, USA) antibodies were used for immunoblotting.

Techniques: Binding Assay, Stable Transfection, Expressing, Transfection, Immunoprecipitation, Western Blot, FLAG-tag, Blue Native PAGE, Electrophoresis, Fluorescence, Confocal Microscopy, Mutagenesis

Journal: Cell Communication and Signaling : CCS

Article Title: Src family kinases interfere with dimerization of STAT5A through a phosphotyrosine-SH2 domain interaction

doi: 10.1186/s12964-014-0081-7

Figure Lengend Snippet: Activated SFK interfere with dimerization and nuclear translocation of pSTAT5A in BCR-ABL expressing cells. Left scheme: Classical activation of the JAK2-STAT5A signaling pathway downstream of the EpoR. Right scheme: BCR-ABL directly phosphorylates STAT5A Y694 resulting in STAT5A dimerization, nuclear accumulation and finally target gene expression . In the presence of BCR-ABL, a predominantly cytoplasmic localization of pSTAT5A is achieved (i) upon binding to the scaffolding adaptor Gab2 resulting in pro-survival signaling through PI3K/Akt activation and (ii) through binding of the STAT5A SH2 domain to the phosphorylated activation loop of SFK, a mechanism that interferes with STAT5A dimerization and subsequent nuclear accumulation. Constitutively active STAT5A S710F escapes the SFK-mediated cytoplasmic retention. Flashes indicate phosphorylation events.

Article Snippet: Anti-pY 694/699 -STAT5A/B (#9351), anti-pY 416 -Src (#2101), anti-pY 412 -Abl (#2865), anti-Hsp70 (#4872, Cell Signaling, Beverly, USA), anti-STAT5A (clone #5073, rabbit polyclonal antiserum was kindly provided by Richard Moriggl, Ludwig Boltzmann Institute for Cancer Research (LBI-CR), Vienna, Austria), anti-GFP (600-103-215, Rockland, Gilbertsville, USA), anti-FLAG (F3165), anti-α-tubulin (T5168, Sigma, St. Louis, USA), anti-GAPDH (sc-32233), anti-Abl (sc-131), anti-Hck (sc-72), anti-cSrc (sc-19, Santa Cruz Biotechnology, Santa Cruz, USA), anti-vSrc (MABS193, Millipore, Billerica, MA, USA) and anti-HA (MMS-101R, Covance, Princeton, New Jersey, USA) antibodies were used for immunoblotting.

Techniques: Translocation Assay, Expressing, Activation Assay, Binding Assay, Scaffolding

Journal: BMC Biology

Article Title: Identification of berberine as a novel drug for the treatment of multiple myeloma via targeting UHRF1

doi: 10.1186/s12915-020-00766-8

Figure Lengend Snippet: BBR induces UHRF1 degradation via the ubiquitin-proteasome system pathway. a BBR affected the stability of UHRF1 in MM cells. RPMI-8266 and MM.1S cells were treated with DMSO alone, BBR (25 μM) alone for 24 h, or pretreated with DMSO or BBR for 12, 16, and 20 h, followed by addition of CHX (50 μg/mL) for additional 4, 8, and 12 h. Cell lysates were harvested and subjected to western blotting with anti-UHRF1 and anti-GAPDH antibodies. b , c Densitometry was utilized to quantify UHRF1 protein levels after normalization with GAPDH control to obtain percent UHRF1 degradation in RPMI-8266 and MM.1S cells. The data were presented as the mean ± SD obtained from three independent experiments. d MG132 abolished the effect of BBR on UHRF1 degradation. RPMI-8266 and MM.1S cells were treated with DMSO alone, BBR (25 μM) alone for 24 h, or pretreated with DMSO or BBR for 20 h, followed by addition of MG132 for additional 4 h. Cell lysates were harvested and subjected to western blotting with anti-UHRF1 and anti-GAPDH antibodies. e RPMI-8266 cells were subsequently treated with BBR (25 μM for 24 h) prior to harvesting. The proteins modified by ubiquitination were purified from cell extracts using anti-UB beads and subjected to western blotting with anti-UHRF1 and anti-GAPDH antibodies. f RPMI-8266 cells were transiently transfected with HA-Ub constructs, and endogenous UHRF1 proteins were immunoprecipitated from BBR-treated or BBR-untreated RPMI-8266 cells (25 μM for 24 h). Immunoprecipitates were harvested and subjected to western blotting with anti-UHRF1, anti-FK2, and anti-GAPDH antibodies

Article Snippet: Ubiquitinated proteins were immunoprecipitated using Signal-SeekerTM Ubiquitination Detection Kit according to the protocol provided by the manufacturer (cat. #BK161, Cytoskeleton).

Techniques: Ubiquitin Proteomics, Western Blot, Control, Modification, Purification, Transfection, Construct, Immunoprecipitation

Journal: Annals of Thoracic Medicine

Article Title: The diagnostic significance of signal peptide-complement C1r/C1s, Uegf, and Bmp1-epidermal growth factor domain-containing protein-1 levels in pulmonary embolism

doi: 10.4103/1817-1737.191876

Figure Lengend Snippet: SCUBE1 levels in PE, non-PE, and control groups. Horizontal lines represent the median of SCUBE1 levels for PE, non-PE, and control groups as 0.90, 0.38, and 0.47 ng/mL, respectively. SCUBE1: Signal peptide-complement C1r/C1s, Uegf, and Bmp1-epidermal growth factor domain-containing protein 1

Article Snippet: Human SCUBE1 ELISA kit (Elabscience Biotechnology Co., Ltd., China, Catalog no: E-EL-H5405, Lot: AK0015NOV30024) was used with BIOTEK semiautomatic ELISA reader.

Techniques: Control

Journal: Annals of Thoracic Medicine

Article Title: The diagnostic significance of signal peptide-complement C1r/C1s, Uegf, and Bmp1-epidermal growth factor domain-containing protein-1 levels in pulmonary embolism

doi: 10.4103/1817-1737.191876

Figure Lengend Snippet: Receiver operating characteristics curve analysis of plasma SCUBE1 values in pulmonary embolism patients. The optimum diagnostic cutoff point of SCUBE1 was 0.49 ng/mL; the area under the curve was 0.791 (95% confidence interval: 0.945–1.000); the sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 64%, 56%, and 100%, respectively. SCUBE1: Signal peptide-complement C1r/C1s, Uegf, and Bmp1-epidermal growth factor domain-containing protein 1

Article Snippet: Human SCUBE1 ELISA kit (Elabscience Biotechnology Co., Ltd., China, Catalog no: E-EL-H5405, Lot: AK0015NOV30024) was used with BIOTEK semiautomatic ELISA reader.

Techniques: Clinical Proteomics, Diagnostic Assay