Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Specific CD8 + T cell responses to HLA-A2 restricted MAGE-A3 p271–279 peptide in hepatocellular carcinoma patients without vaccination

doi: 10.1007/s00262-007-0338-8

Figure Lengend Snippet: Cytotoxic function of M3271 specific CD8+ T cells against MAGE-A3 expressing HLA-A2+ HCC cells. The effector CD8+ T cells from CL085, the sample contains potent CD8+ T cells responsive to M3271 as assessed by IFN-γ release ELISPOT and tetramer assays, were tested for their intracellular perforin expression (a), the Granzyme B- and IFN-γ release after restimulation with MAGE-A3+HLA-A2+ primary HCC cell lines (b, Ch-hep-3, Ch-hep-4 and Ch-hep-6), and the cytotoxicity to the targets of peptide pulsed T2 cells assessed by LDH release assay (c). The CD45 phenotype of effector CD8+ T cells was identified (d). The effector CD8+ T cells of patient GL085 were divided into two groups, cells of group one were stained with PE-tetramer, FITC-anti-CD45RA and perCp-anti-CD8 mAbs, while the cells of group two were stained with PE-tetramer, FITC-anti-CD45RO and perCp-anti-CD8 mAbs. Cells were gated on CD8+ T cells

Article Snippet: Cells were first stained with phycoerythrin (PE)-labeled M3 271 tetramer and control tetramer for 15 min at room temperature, then, the fluorescein isothiocyanate (FITC)-conjugated CD8 mAb (BD Pharmingen, San Diego, CA, USA) was added for further 15 min staining on ice.

Techniques: Expressing, Enzyme-linked Immunospot, Lactate Dehydrogenase Assay, Staining

Journal: Nature Communications

Article Title: N 1 -methyladenosine methylation in tRNA drives liver tumourigenesis by regulating cholesterol metabolism

doi: 10.1038/s41467-021-26718-6

Figure Lengend Snippet: a Overview of immunohistochemistry analysis of endogenous m 1 A levels in a panel of 191 clinically defined human HCC samples. Samples are arranged by grades and then sorted by microvascular invasion (MVI) (gray: “no”; red: “yes”). Paired samples are connected with gray lines. b Representative immunohistochemistry images of endogenous m 1 A levels (brown) in grade III HCC tumor tissue. Scale bar, 100 μm. n = 4 independent experiments. c LC-MS/MS quantification of m 1 A/A and m 6 A/A ratios in total RNA purified from HCC samples. Data are means ± SD. n = 4 biologically independent experiments. Exact P values from left to right: 0.052, 0.00082, 0.00043, 0.026, 0.011, 0.043, 0.0075, 0.13, 0.084, 0.055. d Kaplan–Meier plots of overall survival based on m 1 A levels in the tissue microarray assay (TMA) cohort. The median expression value is used as a cut-off. P values for Kaplan–Meier curves were calculated using a log-rank test. e LC-MS/MS quantification of m 1 A levels in total RNA, large RNA (≥200 nucleotide, nt, mostly composed of rRNA) and small RNA (<200 nt, mostly composed of tRNA) purified from HCC primary CSCs and non-CSCs, presented as percentage of unmodified A. Data are means ± SD. n = 5 biologically independent experiments. Exact P values from left to right: 0.0052, 0.0063, 0.0065, 0.067, 0.11, 0.075, 0.0017, 0.0089, and 0.0084. f Global m 1 A levels were detected in liver CSCs (C) and non-CSCs (N) from HCC primary tumors using dot blot assay. MB (methyl blue) was used as loading controls. n = 4 biologically independent samples. g Representative immunofluorescence staining of m 1 A and CD13 in HCC primary tumor cells and respective oncosphere cells (S). DAPI, 4,6-diamidino-2-phenylindole. Scale bar, 10 μm. n = 4 biologically independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001, and NS, not significant ( P > 0.05) by two-tailed Student’s t -test.

Article Snippet: PE-anti-human IgG (12-4998-82), fluorescein isothiocyanate (FITC)-anti-human CD13 (clone no. WM-15, 11-0138-42), FITC-anti-human IgG (31529) were purchased from eBioscience (San Diego, USA).

Techniques: Immunohistochemistry, Liquid Chromatography with Mass Spectroscopy, Purification, Microarray, Expressing, Dot Blot, Immunofluorescence, Staining, Two Tailed Test

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: GDF15 is an epithelial-derived biomarker of idiopathic pulmonary fibrosis

doi: 10.1152/ajplung.00062.2019

Figure Lengend Snippet: Genes encoding secreted proteins that are transcriptionally upregulated in senescent murine type II alveolar epithelial cells

Article Snippet: Membranes were blocked and incubated with primary antibodies at 4°C overnight from the following sources: GDF15 (G-5, Santa Cruz Biotechnology; recognizes proGDF15), and GAPDH (FL-335, Santa Cruz Biotechnology).

Techniques: Migration

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: GDF15 is an epithelial-derived biomarker of idiopathic pulmonary fibrosis

doi: 10.1152/ajplung.00062.2019

Figure Lengend Snippet: Growth and differentiation factor 15 ( Gdf15 ) is upregulated in response to telomere dysfunction. A : schematic of our analysis strategy for identifying secreted proteins from transcriptional profiling data. Transcriptional data were obtained as described previously . Differentially upregulated genes that were also annotated as secreted proteins are identified. B : quantitative real-time PCR for Gdf15 from sorted type II alveolar epithelial cells (AEC2s) from Trf2 Fl/+ Sftpc-CreER (control) and Trf2 Fl/Fl Sftpc-CreER (senescent) AEC2s. Cells were sorted 10 days after treatment with tamoxifen based on green fluorescent protein expression from mTmG reporter allele . Gene expression was normalized to Hprt and B2m . C : representative images of RNA in situ hybridization staining for Gdf15 from mouse lungs 6 wk after treatment with tamoxifen showing AEC2-specific expression of Gdf15 . AEC2s were identified by expression of the Sftpc transcript. Scale bar is 50 µM. D : quantitation of the colocalization of Gdf15 and Sftpc transcripts ( n = 4 mice per group). E : bronchoalveolar lavage cell counts from mice treated with 2 µg of GDF15. GDF15 or sterile saline was instilled directly into the lungs and the bronchoalveolar lavage was collected thereafter at the indicated times. Total viable cells were quantitated by trypan blue staining, and a differential count was performed on >100 cells. Values are means and standard deviation (SD). Student’s t test (two-tailed) was used to compare groups.

Article Snippet: Membranes were blocked and incubated with primary antibodies at 4°C overnight from the following sources: GDF15 (G-5, Santa Cruz Biotechnology; recognizes proGDF15), and GAPDH (FL-335, Santa Cruz Biotechnology).

Techniques: Real-time Polymerase Chain Reaction, Control, Expressing, Gene Expression, RNA In Situ Hybridization, Staining, Quantitation Assay, Sterility, Saline, Standard Deviation, Two Tailed Test

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: GDF15 is an epithelial-derived biomarker of idiopathic pulmonary fibrosis

doi: 10.1152/ajplung.00062.2019

Figure Lengend Snippet: Bleomycin induces expression of growth and differentiation factor 15 (GDF15). A and B : quantitative ELISA of GDF15 levels in bronchoalveolar lavage (BAL; A ) and plasma ( B ) from mice treated with intratracheal bleomycin or saline. Mice were treated on day 0 , and groups of 6–7 mice (at least 3 male and 3 female at each time point) were harvested at the indicated time points. C : representative RNA in situ hybridization of day 3 lungs showing alveolar expression of Gdf15 . D : quantitation of colocalization of Sftpc and Gdf15 transcripts in RNA in situ hybridization staining. Values are means and SD. Student’s t test (two-tailed) was used to compare groups. **** P < 0.0001, *** P < 0.001, and ** P < 0.01.

Article Snippet: Membranes were blocked and incubated with primary antibodies at 4°C overnight from the following sources: GDF15 (G-5, Santa Cruz Biotechnology; recognizes proGDF15), and GAPDH (FL-335, Santa Cruz Biotechnology).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Saline, RNA In Situ Hybridization, Quantitation Assay, Staining, Two Tailed Test

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: GDF15 is an epithelial-derived biomarker of idiopathic pulmonary fibrosis

doi: 10.1152/ajplung.00062.2019

Figure Lengend Snippet: Growth and differentiation factor 15 (GDF15) is expressed by honeycomb cyst epithelial cells. A : representative photomicrographs from three independent donor and idiopathic pulmonary fibrosis (IPF) lungs. Slides were stained for GDF15 (brown) and counterstained with hematoxylin. GDF15 expression is present in macrophages from healthy lungs (arrowheads) but rarely in epithelial cells. In contrast, GDF15 expression was abundant in epithelial cells (arrows) and macrophages in fibrotic lungs. Scale bar in micrographs is 2 mm and 100 μm in insets . B : GDF15 RNA in situ hybridization (RNA-ISH) in donor and IPF lungs showing epithelial-specific expression of GDF15 . Scale bar is 100 μm. C : Western blot of whole lung lysate from donor and IPF lungs for proGDF15 and GAPDH as a load control. D : quantitation of proGDF15 in Western blot in C . Values are means and SD. Student’s t test, two tailed was used for comparison in D . IHC, immunohistochemistry.

Article Snippet: Membranes were blocked and incubated with primary antibodies at 4°C overnight from the following sources: GDF15 (G-5, Santa Cruz Biotechnology; recognizes proGDF15), and GAPDH (FL-335, Santa Cruz Biotechnology).

Techniques: Staining, Expressing, RNA In Situ Hybridization, Western Blot, Control, Quantitation Assay, Two Tailed Test, Comparison, Immunohistochemistry

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: GDF15 is an epithelial-derived biomarker of idiopathic pulmonary fibrosis

doi: 10.1152/ajplung.00062.2019

Figure Lengend Snippet: Growth and differentiation factor 15 (GDF15) is a biomarker of idiopathic pulmonary fibrosis (IPF) and identifies high-risk patients. A–C : plasma levels of GDF15 in three cohorts of controls and IPF patients. GDF15 was measured using Luminex assay in cohort 1 ( A ) and ELISA in cohorts 2 and 3 ( B and C , respectively). Values are means and SD. D and E : Kaplan-Meier graph showing the proportion of patients who were alive or transplant free as a function of time. IPF patients were split into two groups based on GDF15 level. Patients from the upper quartile were compared with the lower three quartiles. Age-adjusted P values are from Cox regression analysis. Comparisons in A , B , and C used Welch’s t test, two tailed.

Article Snippet: Membranes were blocked and incubated with primary antibodies at 4°C overnight from the following sources: GDF15 (G-5, Santa Cruz Biotechnology; recognizes proGDF15), and GAPDH (FL-335, Santa Cruz Biotechnology).

Techniques: Biomarker Discovery, Clinical Proteomics, Luminex, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: GDF15 is an epithelial-derived biomarker of idiopathic pulmonary fibrosis

doi: 10.1152/ajplung.00062.2019

Figure Lengend Snippet: Clinical characteristics and correlation between plasma GDF15 and pulmonary function in patients with IPF

Article Snippet: Membranes were blocked and incubated with primary antibodies at 4°C overnight from the following sources: GDF15 (G-5, Santa Cruz Biotechnology; recognizes proGDF15), and GAPDH (FL-335, Santa Cruz Biotechnology).

Techniques: Clinical Proteomics, Luminex, Enzyme-linked Immunosorbent Assay