Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: RETRACTED: Blockade of TNF-α signaling suppresses the AREG-mediated IL-6 and IL-8 cytokines secretion induced by anti-Ro/SSA autoantibodies.

doi: 10.1038/labinvest.2010.168

Figure Lengend Snippet: Figure 1 Semiquantitative RT-PCR and real-time PCR for Furin, TACE and AREG genes expression. (a) RT-PCR analysis of Furin, TACE and AREG mRNA extracted from SGEC treated with anti-Ro/SSA autoantibodies. M, marker; control, untreated SGEC; HIgG, SGEC treated with IgG fractions extracted from sera of healthy donors; anti-Ro, SGEC treated with anti-Ro/SSA autoantibodies. RT-PCR of GADPH was used as control. Band intensities were analyzed by densitometry (b). (c) Real-time PCR for Furin, TACE and AREG genes expression. Representative histograms of mRNA levels of Furin, TACE and AREG in untreated SGEC (control), SGEC treated with healthy IgG (HIgG), anti-Ro/SSA autoantibodies (anti-Ro). The mRNA levels of the housekeeping gene, b-2 microglobulin, were quantified between untreated control cells and variously treated cells (data represent the mean±s.e. of five independent experiments).

Article Snippet: Membranes were incubated for 90 min with rabbit anti-human Furin polyclonal antibody (pAb), goat antihuman TACE pAb (both from Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse anti-human AREG mAb (R&D Systems, Minneapolis, MN USA), and for 30 min with the relative secondary antibodies-HRP conjugates (Santa Cruz Biotechnology).

Techniques: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Expressing, Marker, Control

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: RETRACTED: Blockade of TNF-α signaling suppresses the AREG-mediated IL-6 and IL-8 cytokines secretion induced by anti-Ro/SSA autoantibodies.

doi: 10.1038/labinvest.2010.168

Figure Lengend Snippet: Figure 2 Analysis of Furin, TACE and AREG expression in anti-Ro/SSA Abs-treated SGEC. (a) Flow cytometric analysis of Furin, TACE and AREG expression in SGEC after anti-Ro/SSA Abs treatment. Examples of flow cytometric images from one representative experiment. (A) Furin expression analysis in untreated and anti-Ro/SSA or HIgG-treated SGEC; (B) intracellular active TACE expression analysis in untreated and anti-Ro/SSA or HIgG-treated SGEC; (C) AREG expression analysis in untreated and anti-Ro/SSA or HIgG-treated SGEC. (b) Western blot analysis of Furin, TACE and AREG proteins expression in SGEC treated or not with anti-Ro/SSA. Immunoblotting gave rise to bands of the expected size (97 kDa for Furin, 80 kDa for active TACE and 50 kDa for AREG). b-Actin was used as protein loading control. (c) Detection of soluble AREG by ELISA. Secreted AREG was detected by ELISA in the conditioned medium. Control, untreated SGEC; HIgG, SGEC treated with IgG fractions extracted from sera of healthy donors; anti-Ro, SGEC treated with anti-Ro/SSA autoantibodies. (Data represent the mean±s.e. of four independent experiments).

Article Snippet: Membranes were incubated for 90 min with rabbit anti-human Furin polyclonal antibody (pAb), goat antihuman TACE pAb (both from Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse anti-human AREG mAb (R&D Systems, Minneapolis, MN USA), and for 30 min with the relative secondary antibodies-HRP conjugates (Santa Cruz Biotechnology).

Techniques: Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Orally Delivered Milk-Derived Nanovesicles Loaded with Connexin 43 Peptides for Targeted Cardiac Ischemia-Reperfusion Therapy

doi: 10.1101/2025.01.01.630994

Figure Lengend Snippet: A) Schematic of mEV isolation protocol showing the steps of the procedure. Fractions referred to are those collected during Sepharose Column (SEC) fractionation. Fractions 7 through 9 (F7-F9) show highest levels of mEV enrichment and samples used in this study were pooled from F7-F9. Fractions >14 show enrichment for contaminating milk proteins such as casein, with low or no evidence of mEVs, and are typically discarded. B) Single Molecule Localization Microscopy (SMLM) image of an isolated mEV isolated by the protocol in (A) tagged for CD81 (red) and CD63 (teal). C) Negative stain transmission electron microscopy (TEM) indicating dense accumulations of mEVs generated by the isolation protocol. D) Calcein AM loading assay illustrating membrane intactness of isolated mEVs E) Western blotting of mEV isolates for CD81, CD9, TSG-101, and negativity and markedly reduced expression of Calnexin and Casein, respectively. Lysates from HeLa cells are included as a control. Sepharose fraction 16 (F16) shows elevated casein, but no evidence of EV markers. F) Nanoparticle Tracking Analysis (NTA) of an mEV isolate indicating >5×10 12 mEVs per mL, with average nanoparticle size of 125.0 nm, consistent with small EVs. G) SMLM-generated size distribution of CD9/CD81/CD63-positive mEVs, showing a similar histogram morphology to NTA results in panel F.

Article Snippet: Overnight primary antibody incubation was performed and primary antibodies were diluted in the blocking buffer as follows: CD81 (Cell Signaling Technology, 56039S, 1:1000) CD9 (Novus Biologicals, NB500-494, 1:1000), Calnexin (MilliporeSigma, AB2301, 1:5000), TSG101 (Bethyl Laboratories Inc. A303-506A, 1:5000), Casein (Abcam, Ab166596, 1:2000), Cx43 Cytoplasmic-Loop (Invitrogen, PA5-11632, 1:1000), and antibodies against the Cx43 CT (MilliporeSigma, C6219, 1:4000 and Abcam, Ab87645, 1:1000).

Techniques: Isolation, Fractionation, Microscopy, Staining, Transmission Assay, Electron Microscopy, Generated, Membrane, Western Blot, Expressing, Control

Journal: Oncotarget

Article Title: Down-regulation of CITED2 attenuates breast tumor growth, vessel formation and TGF-β-induced expression of VEGFA

doi: 10.18632/oncotarget.14048

Figure Lengend Snippet: ( A ) Average tumor volume measured over time following injection of scramble or shCITED2-expressing MDA-MB-231 (left) and MDA-MB-468 (right) cells into the mammary fat pad of athymic nude mice. ( B ) Representative H&E-stained sections of tumor from each experimental group. The pale pink areas highlighted by yellow dotted lines denote deceased tissues. The adjacent histogram represents the average percentage of deceased tissue displayed by the tumors in each experimental group as measured on H&E-stained sections. ( C ) Representative images of immunohistochemical analysis of Ki67 protein expression from each experimental group. Ki67 was identified using DAB (brown) and visualized by light microscopy. The adjacent histogram represents the average percentage of Ki67 positive cells displayed in each experimental group. (** P < 0.01, *** P < 0.001). Scale bar: 4 mm.

Article Snippet: Endogenous biotin was then blocked using an Avidin and Biotin blocking kit (DAKO) for 10 minutes and incubated at 37°C for one hour with rabbit anti-human ki67 antibody (1:20 dilution, NB500-0170, Novus Biologicals).

Techniques: Injection, Expressing, Staining, Immunohistochemical staining, Light Microscopy

Journal: British Journal of Cancer

Article Title: Expression of cytoplasmic and nuclear Survivin in primary and secondary human glioblastoma

doi: 10.1038/sj.bjc.6602904

Figure Lengend Snippet: Immunohistochemical staining of Survivin and fluorescent TUNEL staining in human gliomas ( A–G ). A primary glioblastoma showed a high level (++++) expression of nuclear Survivin ( A ). High staining score (++++) of cytoplasmic Survivin was detected in another primary glioblastoma ( B ). Moderate expression level (++) of cytoplasmic Survivin was detected in a pre-existing low-graded (grade II) glioma ( C ), which was obviously lower than that in its paired secondary glioblastoma with a staining score of ++++ ( D ). High-level (++++) expression of nuclear Survivin was detected in a secondary glioblastoma ( E ) and its matched pre-existing grade II glioma ( F ). Three apoptotic cells with clear nuclear staining (green color) were examined in a primary glioblastoma ( G ).

Article Snippet: The tumour slides were incubated at 4°C overnight in a moist chamber with one of the two polyclonal anti-Survivin antibodies (NB-500-201 K3, Novus Biologicals, Littleton, CO, USA, 1 : 300 dilution and SC-10811, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 400 dilution).

Techniques: Immunohistochemical staining, Staining, TUNEL Assay, Expressing

Journal: British Journal of Cancer

Article Title: Expression of cytoplasmic and nuclear Survivin in primary and secondary human glioblastoma

doi: 10.1038/sj.bjc.6602904

Figure Lengend Snippet: Status of Survivin expression and apoptotic index in primary GBMs

Article Snippet: The tumour slides were incubated at 4°C overnight in a moist chamber with one of the two polyclonal anti-Survivin antibodies (NB-500-201 K3, Novus Biologicals, Littleton, CO, USA, 1 : 300 dilution and SC-10811, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 400 dilution).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Expression of cytoplasmic and nuclear Survivin in primary and secondary human glioblastoma

doi: 10.1038/sj.bjc.6602904

Figure Lengend Snippet: Status of Survivin expression and apoptotic index in secondary GBMs

Article Snippet: The tumour slides were incubated at 4°C overnight in a moist chamber with one of the two polyclonal anti-Survivin antibodies (NB-500-201 K3, Novus Biologicals, Littleton, CO, USA, 1 : 300 dilution and SC-10811, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 400 dilution).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Expression of cytoplasmic and nuclear Survivin in primary and secondary human glioblastoma

doi: 10.1038/sj.bjc.6602904

Figure Lengend Snippet: The expression of cytoplasmic and nuclear Survivin in 15 paired secondary GBMs and pre-existing lesions a

Article Snippet: The tumour slides were incubated at 4°C overnight in a moist chamber with one of the two polyclonal anti-Survivin antibodies (NB-500-201 K3, Novus Biologicals, Littleton, CO, USA, 1 : 300 dilution and SC-10811, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 400 dilution).

Techniques: Expressing