Journal: Stem Cell Research & Therapy

Article Title: Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy

doi: 10.1186/s13287-016-0463-4

Figure Lengend Snippet: Human umbilical cord-derived mesenchymal stem cell exosomes ( hucMSC-Ex ) prevent deterioration of renal function in vivo. a Representative images of renal histology (200×, scale bar = 50 μm). b The histomorphological score. c Immunohistochemical analysis of PCNA expression in kidney tissues (200×, scale bar = 50 μm). d Percentage of proliferating cell nuclear antigen ( PCNA )-positive cells. e TUNEL assay for apoptotic cells in rat renal tissues (200×, scale bar = 50 μm). f Percentage of apoptotic cells. Serum creatinine ( Cr ) ( g ) and blood urea nitrogen ( BUN ) ( h ) levels were measured at different times after cisplatin injection. Data are expressed as mean ± SD. * P < 0.05, n = 6. Rats treated as mentioned for the in vivo rat model were divided into four groups as follows: 1) control; 2) phosphate-buffered saline ( PBS ); 3) hucMSC-Ex; and 4) human fetal lung fibroblast-1exosomes ( HFL1-Ex ). All animals were sacrificed at 3 days after cisplatin injection

Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was used to detect the apoptotic cells. hucMSCs, HFL1 cells, NRK-52E cells, and the kidney tissues were examined using the TdT-FragEL DNA fragmentation detection kit according to the manufacturer's protocol (Boster, China).

Techniques: Derivative Assay, In Vivo, Immunohistochemical staining, Expressing, TUNEL Assay, Injection, Control, Saline

Journal: Stem Cell Research & Therapy

Article Title: Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy

doi: 10.1186/s13287-016-0463-4

Figure Lengend Snippet: Human umbilical cord-derived mesenchymal stem cell exosomes ( hucMSC-Ex ) inhibit cisplatin-induced apoptosis and secretion of inflammatory cytokines in NRK-52E cells in vitro. a NRK-52E cells were stained with JC-1 dye and observed under fluorescent microscopy (JC-1 produces red fluorescence within the mitochondria as JC-1-aggregates while it emits green fluorescence when it leaks into the cytoplasm as JC-1-monomers; the shift between green and red is proportional to the mitochondrial membrane potential change) (200×, scale bar = 50 μm). b TUNEL assay was performed to detect the apoptotic cells (200×, scale bar = 50 μm). c Percentage of apoptotic cells. ** P < 0.01, n = 3. d The expression of proliferating cell nuclear antigen ( PCNA ), BCL-XL, BCL-2, Bax, Cytochrome-c, cleaved Caspase-3, and GAPDH proteins was evaluated by Western blot. e Immunohistochemical analysis of tumor necrosis factor alpha ( TNF-α ) expression (200×, scale bar = 50 μm). f The expression of interleukin 1-β ( IL1-β ) and GAPDH proteins was evaluated by Western blot. NRK-52E cells treated as mentioned for the in vitro experiment were divided into four groups: 1) control; 2) phosphate-buffered saline ( PBS ); 3) hucMSC-Ex; and 4) human fetal lung fibroblast-1exosomes ( HFL1-Ex )

Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was used to detect the apoptotic cells. hucMSCs, HFL1 cells, NRK-52E cells, and the kidney tissues were examined using the TdT-FragEL DNA fragmentation detection kit according to the manufacturer's protocol (Boster, China).

Techniques: Derivative Assay, In Vitro, Staining, Microscopy, Fluorescence, Membrane, TUNEL Assay, Expressing, Western Blot, Immunohistochemical staining, Control, Saline

Journal: Stem Cell Research & Therapy

Article Title: Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy

doi: 10.1186/s13287-016-0463-4

Figure Lengend Snippet: Human umbilical cord-derived mesenchymal stem cell exosomes ( hucMSC-Ex ) prevent apoptosis and secretion of inflammatory cytokines by activating autophagy in vivo. a Representative images of renal histology (200×, scale bar = 50 μm). b The histomorphological score. c Immunohistochemical analysis of PCNA expression in kidney tissues (200×, scale bar = 50 μm). d Percentage of proliferating cell nuclear antigen ( PCNA )-positive cells. e TUNEL assay for apoptotic cells in rat renal tissues (200×, scale bar = 50 μm). f Percentage of apoptotic cells. g The expression of LC3B, BCL-2, Bax, IL1-β, and GAPDH proteins in the cisplatin-induced rat model were measured by Western blot. The serum inflammatory cytokines ( h ) tumor necrosis factor alpha ( TNF-α ), ( i ) interleukin-1β ( IL1-β ), and ( j ) interleukin-6 ( IL6 ) were measured via Luminex assay. Data are expressed as mean ± SD. * P < 0.05, n = 6. Rats were divided into six groups as follows: 1) control; 2) phosphate-buffered saline ( PBS ); 3) hucMSC-Ex; 4) hucMSC-Ex + 3-methyladenine ( 3MA ); 5) 3MA; and 6) rapamycin ( Rapa )

Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was used to detect the apoptotic cells. hucMSCs, HFL1 cells, NRK-52E cells, and the kidney tissues were examined using the TdT-FragEL DNA fragmentation detection kit according to the manufacturer's protocol (Boster, China).

Techniques: Derivative Assay, In Vivo, Immunohistochemical staining, Expressing, TUNEL Assay, Western Blot, Luminex, Control, Saline

Journal: Translational Oncology

Article Title: Proteomic analysis reveals distinctive protein expression patterns of thrombus in clear cell renal cell carcinoma

doi: 10.1016/j.tranon.2020.100895

Figure Lengend Snippet: The GO enrichment results of the proteins in cluster 1–5.

Article Snippet: Slides were reacted for 2 h at room temperature with primary rabbit polyclonal antibody to CFHR1 (Abcam; Rabbit; 1:500 dilution), KRT7 (Bioworld; Rabbit; 1:500 dilution), GGT5 (Bioworld; Rabbit; 1:500 dilution).

Techniques: Binding Assay, Migration, Membrane, Activity Assay, Protein Binding, Clinical Proteomics

Journal: Translational Oncology

Article Title: Proteomic analysis reveals distinctive protein expression patterns of thrombus in clear cell renal cell carcinoma

doi: 10.1016/j.tranon.2020.100895

Figure Lengend Snippet: Western blot and immunohistochemistry analysis of CFHR1, GGT5 and KRT7. (A) Examples for the western blot result of CFHR1, GGT5 and KRT7 from patient #5. (B) Statistical result of the western blot analysis of the three proteins in the validation cohort comprised of 15 ccRCC patients. The amount of target proteins are calculated by gray scanning and presented as column graph. (C) H&E staining and immunohistochemistry analysis result of the three proteins in thrombus, tumor and normal tissue. Group labels used: N, normal tissue; C, tumor; T, thrombus.

Article Snippet: Slides were reacted for 2 h at room temperature with primary rabbit polyclonal antibody to CFHR1 (Abcam; Rabbit; 1:500 dilution), KRT7 (Bioworld; Rabbit; 1:500 dilution), GGT5 (Bioworld; Rabbit; 1:500 dilution).

Techniques: Western Blot, Immunohistochemistry, Biomarker Discovery, Staining

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Complement Proteins’ Western Blotting Information.

Article Snippet: 9 , CFHR1 , 37 kDa , Mouse anti-Factor H-related 1 (CFHR1) Monoclonal antibody #MAB4247 (RD Systems) , 1:1000 , Human , Anti-mouse IgG, HRP-linked antibody #7076 (CST) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Complement Proteins’ Western Blotting Information.

Article Snippet: 9 , CFHR1 , 37 kDa , Mouse anti-Factor H-related 1 (CFHR1) Monoclonal antibody #MAB4247 (RD Systems) , 1:1000 , Human , Anti-mouse IgG, HRP-linked antibody #7076 (CST) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Western Blot

Journal: Translational Oncology

Article Title: Proteomic analysis reveals distinctive protein expression patterns of thrombus in clear cell renal cell carcinoma

doi: 10.1016/j.tranon.2020.100895

Figure Lengend Snippet: The GO enrichment results of the proteins in cluster 1–5.

Article Snippet: The proteins were then transferred to a PVDF membrane and probed with primary antibodies: anti-CFHR1 (1,1000; Abcam, Cambridge, MA), anti-KRT7 (1,1000; Bioworld Technology, Minneapolis, MN), anti-GGT5 (1,1000; Bioworld Technology, Minneapolis, MN, USA), and anti-β-actin (1,1000; Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Binding Assay, Migration, Activity Assay, Protein Binding

Journal: Translational Oncology

Article Title: Proteomic analysis reveals distinctive protein expression patterns of thrombus in clear cell renal cell carcinoma

doi: 10.1016/j.tranon.2020.100895

Figure Lengend Snippet: Western blot and immunohistochemistry analysis of CFHR1, GGT5 and KRT7. (A) Examples for the western blot result of CFHR1, GGT5 and KRT7 from patient #5. (B) Statistical result of the western blot analysis of the three proteins in the validation cohort comprised of 15 ccRCC patients. The amount of target proteins are calculated by gray scanning and presented as column graph. (C) H&E staining and immunohistochemistry analysis result of the three proteins in thrombus, tumor and normal tissue. Group labels used: N, normal tissue; C, tumor; T, thrombus.

Article Snippet: The proteins were then transferred to a PVDF membrane and probed with primary antibodies: anti-CFHR1 (1,1000; Abcam, Cambridge, MA), anti-KRT7 (1,1000; Bioworld Technology, Minneapolis, MN), anti-GGT5 (1,1000; Bioworld Technology, Minneapolis, MN, USA), and anti-β-actin (1,1000; Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Western Blot, Immunohistochemistry, Staining

Journal: Translational Oncology

Article Title: Proteomic analysis reveals distinctive protein expression patterns of thrombus in clear cell renal cell carcinoma

doi: 10.1016/j.tranon.2020.100895

Figure Lengend Snippet: The GO enrichment results of the proteins in cluster 1–5.

Article Snippet: The proteins were then transferred to a PVDF membrane and probed with primary antibodies: anti-CFHR1 (1,1000; Abcam, Cambridge, MA), anti-KRT7 (1,1000; Bioworld Technology, Minneapolis, MN), anti-GGT5 (1,1000; Bioworld Technology, Minneapolis, MN, USA), and anti-β-actin (1,1000; Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Binding Assay, Migration, Activity Assay, Protein Binding

Journal: Translational Oncology

Article Title: Proteomic analysis reveals distinctive protein expression patterns of thrombus in clear cell renal cell carcinoma

doi: 10.1016/j.tranon.2020.100895

Figure Lengend Snippet: Western blot and immunohistochemistry analysis of CFHR1, GGT5 and KRT7. (A) Examples for the western blot result of CFHR1, GGT5 and KRT7 from patient #5. (B) Statistical result of the western blot analysis of the three proteins in the validation cohort comprised of 15 ccRCC patients. The amount of target proteins are calculated by gray scanning and presented as column graph. (C) H&E staining and immunohistochemistry analysis result of the three proteins in thrombus, tumor and normal tissue. Group labels used: N, normal tissue; C, tumor; T, thrombus.

Article Snippet: The proteins were then transferred to a PVDF membrane and probed with primary antibodies: anti-CFHR1 (1,1000; Abcam, Cambridge, MA), anti-KRT7 (1,1000; Bioworld Technology, Minneapolis, MN), anti-GGT5 (1,1000; Bioworld Technology, Minneapolis, MN, USA), and anti-β-actin (1,1000; Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Western Blot, Immunohistochemistry, Staining

Journal: Frontiers in Immunology

Article Title: A Family Affair: Addressing the Challenges of Factor H and the Related Proteins

doi: 10.3389/fimmu.2021.660194

Figure Lengend Snippet: Published antibodies that have been proposed to be specific for each of the Factor H-related proteins.

Article Snippet: Mouse mAb anti-human FHR-1 , - IHC on material of patients with a homozygous CFHR-1 deficiency. , IHC , (#3078-M01; Abnova, Taipei, Taiwan) , ( , ) .

Techniques: Biomarker Discovery, Direct ELISA, Double Staining, Immunohistochemistry, Recombinant, Enzyme-linked Immunosorbent Assay, Immunohistochemistry-IF, Mass Spectrometry, Immunoprecipitation, Western Blot

Journal: Clinical and Developmental Immunology

Article Title: Contribution of the Infection-Associated Complement Regulator-Acquiring Surface Protein 4 (ErpC) to Complement Resistance of Borrelia burgdorferi

doi: 10.1155/2012/349657

Figure Lengend Snippet: Identification of serum proteins that bind to recombinant CRASP-4. Recombinant, polyhistidine-tagged CRASP-4 was immobilized onto magnetic beads and incubated with NHS. Uncoated beads were also treated under the same conditions and used as a control to identify nonspecific binding of serum proteins. After extensive washing, bound proteins were eluted with 100mM glycine-HCl (pH 2.0) and the eluate fractions were separated by SDS-PAGE under nonreducing conditions. (a) Silver stain of a gel loaded with purified polyhistidine-tagged CRASP-4 (1 μ g), eluate fraction of the uncoated beads, and the final wash and eluate fraction of CRASP-4-coated beads. (b) Western blot analysis of the eluate fraction of CRASP-4-coated beads using a polyclonal anti-CFH or a polyclonal anti-CFHR1 antiserum. Mobilities of molecular mass standards are indicated to the left.

Article Snippet: A polyclonal anti-CFH antiserum was utilized to detect human CFH, CFHR1, and CFHR2 (Merck Biosciences, Bad Soden, Germany and Complement Technology, Tyler, TE).

Techniques: Recombinant, Magnetic Beads, Incubation, Control, Binding Assay, SDS Page, Silver Staining, Purification, Western Blot

Journal: Clinical and Developmental Immunology

Article Title: Contribution of the Infection-Associated Complement Regulator-Acquiring Surface Protein 4 (ErpC) to Complement Resistance of Borrelia burgdorferi

doi: 10.1155/2012/349657

Figure Lengend Snippet: CRASP-4 binds distinct complement proteins. Binding of equimolar amounts of CFH, CFHR1, CFHR2, and CFHR5 (33 μ M) to immobilized CRASP-4 (5 μ g/mL) was analyzed by ELISA. Bound CFH or CFHR proteins were detected with either goat CFH polyclonal antiserum or mouse CFHR1 monoclonal antiserum (JHD 7.10), which reacts with all three CFHRs. Data represent the means and standard errors from three separate experiments.

Article Snippet: A polyclonal anti-CFH antiserum was utilized to detect human CFH, CFHR1, and CFHR2 (Merck Biosciences, Bad Soden, Germany and Complement Technology, Tyler, TE).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Clinical and Developmental Immunology

Article Title: Contribution of the Infection-Associated Complement Regulator-Acquiring Surface Protein 4 (ErpC) to Complement Resistance of Borrelia burgdorferi

doi: 10.1155/2012/349657

Figure Lengend Snippet: Characterization of B. garinii G1 producing CRASP-4. (a) B. garinii G1 and transformed strains G1/pKFSS1 and G1/pCRASP-4 were characterized by PCR amplification using flaB- , aadA- , and erpC -specific primers, as listed in . (b) Synthesis of CRASP-4 by transformed G1 was assessed using ligand affinity blotting. Whole cell lysates (15 μ g each) of G1, G1/pKFSS1 and G1/pCRASP-4 were separated by SDS-PAGE, and transferred to nitrocellulose. After incubation with NHS, binding of CFH to CRASP-4 was identified using a polyclonal antiserum. A monoclonal antibody, L41 1C11, specific for the flagellin protein FlaB, was applied to show equal loading of borrelial lysates. (c) Surface localization of CRASP-4 in transformed G1 cells. Spirochetes were incubated with or without proteinase K or trypsin, respectively, then lysed by sonication, and total proteins were separated by SDS-PAGE. CRASP-4 was identified by ligand affinity analysis as described above. Flagellin (FlaB) was detected with MAb L41 1C11 (dilution 1/1000) by Western blotting. (d) Demonstration of surface expression of CRASP-4 by transformed B. garinii G1, by indirect immunofluoresecence microscopy of intact borrelial cells. Spirochetes were incubated with rabbit polyclonal anti-ErpA/ErpC antiserum before fixation. Periplasmic FlaB, used as control, was detected by mAb L41 1C11 using fixed and unfixed cells. For counterstaining, the DNA-binding dye DAPI was used to identify all bacteria. Slides were visualized at a magnification of ×1,000 using an Olympus CX40 fluorescence microscope mounted with a DS-5Mc charge-coupled device camera (Nikon).

Article Snippet: A polyclonal anti-CFH antiserum was utilized to detect human CFH, CFHR1, and CFHR2 (Merck Biosciences, Bad Soden, Germany and Complement Technology, Tyler, TE).

Techniques: Transformation Assay, Amplification, SDS Page, Incubation, Binding Assay, Sonication, Western Blot, Expressing, Microscopy, Control, Bacteria, Fluorescence

Journal: Clinical and Developmental Immunology

Article Title: Contribution of the Infection-Associated Complement Regulator-Acquiring Surface Protein 4 (ErpC) to Complement Resistance of Borrelia burgdorferi

doi: 10.1155/2012/349657

Figure Lengend Snippet: Binding of serum molecules by B. garinii transformants. B. garinii strains G1, G1/pKFSS1, and G1/pCRASP-4 and B. burgdorferi strain LW2 (used as control) were incubated in NHS plus EDTA to prevent complement activation and washed extensively, and bound proteins were eluted using 0.1 M glycine (pH 2.0). Both the last wash (w) and the eluate (e) fractions obtained from each strain were separated by SDS-PAGE and transferred to nitrocellulose. As an additional control purified CFH (1 μ g) was also applied. Membranes were probed with a polyclonal anti-FHR1 antiserum which recognizes CFH, CFHR1, CFHR2, and CFHR5. Mobilities of molecular mass standards are shown to the left of the panels.

Article Snippet: A polyclonal anti-CFH antiserum was utilized to detect human CFH, CFHR1, and CFHR2 (Merck Biosciences, Bad Soden, Germany and Complement Technology, Tyler, TE).

Techniques: Binding Assay, Control, Incubation, Activation Assay, SDS Page, Purification