Journal: Human Cell

Article Title: Hsa_circ_0000437 promotes the progression of rheumatic valvular heart disease by activating the mitogen-activated protein kinase signaling pathways after sponging let-7f-5p and targeting RAS-like proto-oncogene B

doi: 10.1007/s13577-025-01331-7

Figure Lengend Snippet: hsa_circ_0000437/let-7f-5p/RALB axis promotes inflammatory transformation during RVHD progression by activating the MAPK signaling pathway. Western blotting assay of MAPK14, ERK, JNK, and RALB proteins in hVICs cells transfected with si-hsa_circ_0000437 ( A ), pcDNA3.1-hsa_circ_0000437 ( B ), let-7f-5p mimic and pcDNA3.1-hsa_circ_0000437 ( C ), si-RALB, pcDNA3.1-hsa_circ_0000437 and let-7f-5p inhibitor ( D ). Numbers indicate the relative expression of proteins, and GAPDH serves as an internal reference. E Diagram of the regulatory mechanism of hsa_circ_0000437/let-7f-5p/RALB axis promoting the occurrence and development of RVHD, the target hsa_circ_0000437 was identified by microarray and confirmed in the cytoplasm that RALB could promote inflammatory transformation during RVHD by activating MAPK signaling pathway. RALB was also inhibited by let-7f-5p and enhanced by ceRNA hsa_circ_0000437 (some elements were created using BioRender.com). n = 3, n stands for biological replicates

Article Snippet: MAPK14 (Cat No. 14064–1-AP), extracellular regulated protein kinases (ERK) (Cat No. 11257–1-AP), c-Jun N-terminal kinase (JNK) (Cat No. 51153–1-AP), RAS-like proto-oncogene B (RALB) (Cat No. 12340–1-AP), and goat anti-rabbit IgG and goat anti-mouse IgG secondary antibody were purchased from a subsidiary of Proteintech Wuhan Sanying Biotechnology Co., LTD (China).

Techniques: Transformation Assay, Western Blot, Transfection, Expressing, Microarray

Journal: Frontiers in Pharmacology

Article Title: A senescence-associated signature refines the classification of different modification patterns and characterization of tumor immune microenvironment infiltration in triple-negative breast cancer

doi: 10.3389/fphar.2023.1191910

Figure Lengend Snippet: The hub gene FAM3b and its prognostic value in TNBC. (A) Number of trees showing the importance proportion of SASP regulator genes. (B) FAM3B expression in paired tumor and normal tissues in BRCA from the TCGA database. (C) FAM3B expression according to the molecular subtypes of breast cancer. (D) UMAP plot of intratumoral immune cells and FAM3B showing the correlation between infiltration of different immune cells and FAM3B expression in the Alex, EMTAB8107, GSE150660 and GSE161529 datasets. (E,F) The impact of FAM3B on OS and DFS in breast cancer tissue microarray using Kaplan-Meier analysis.

Article Snippet: IHC staining of FAM3B protein expression in the tissue microarray was performed by incubation with rabbit polyclonal antibodies against human FAM3B antibody (27131-1-AP, Proteintech, 1:200) overnight, followed by incubation with goat monoclonal antibody against rabbit antibody (111-035-003, JACKSON, 1:1,000) for 1 h at room temperature.

Techniques: Expressing, Microarray

Journal: Stem Cell Research & Therapy

Article Title: Oxidant therapy improves adipogenic differentiation of adipose-derived stem cells in human wound healing

doi: 10.1186/s13287-021-02336-3

Figure Lengend Snippet: Characterization of the macrophage activation model. a THP1 cell treatment protocol for in vitro polarization of macrophages. b Morphology (phase contrast, scale bar 100μm) of THP1-derived macrophages on day of CM harvest. c Heatmap of log2 cytokine protein levels in differentially activated macrophages. The dendrogram represents the result from a complete linkage hierarchical clustering of mean-centered expression levels based on Euclidian similarity. d mRNA-expression levels of macrophage-selective markers (M (IFNG/LPS) : CD80, IL1B, and M (IL4/IL13) : CD200R, TGM2). Data were normalized to the 18S rRNA reference gene and shown as mean –ΔCt. Asterisks indicate p values < 0.05 (*), p <0.01 (**), and p <0.001 (***). Statistical significance ( n =5) was determined by using one-way ANOVA and Dunnett’s test for multiple comparison

Article Snippet: Macrophage-CM was loaded onto RayBio® Human Cytokine G5 Antibody Microarray Glass Chip (RayBiotech, Norcross, GA) which facilitates the detection of 80 targets.

Techniques: Activation Assay, In Vitro, Derivative Assay, Expressing

Journal: Mediators of Inflammation

Article Title: LR12 Promotes Liver Repair by Improving the Resolution of Inflammation and Liver Regeneration in Mice with Thioacetamide- (TAA-) Induced Acute Liver Failure

doi: 10.1155/2021/2327721

Figure Lengend Snippet: LR12 promoted hepatocyte regeneration via CCL20 secreted by macrophages. (a) The cytokine protein microarray of the supernatant from macrophages. (b) ELISA of CCL20 in the supernatant from macrophages. (c) The mRNA level of CCL20 in macrophages. (d) CLSM showing F4/80 and CCL20 staining in the liver tissues. (e) Western blot analysis of PCNA in LO2 cells stimulated with CCL20. Data were presented as the mean ± standard deviation (SD). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 versus control group.

Article Snippet: The RayBio Human Cytokine Antibody Array Kit (QAH-TH17-1-1 microarray; RayBiotech, Norcross, GA, USA) was utilized to identify the cytokines based on instructions provided by the manufacturer.

Techniques: Microarray, Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Standard Deviation

Journal: Cell Death & Disease

Article Title: Single-cell sequencing resolves the landscape of immune cells and regulatory mechanisms in HIV-infected immune non-responders

doi: 10.1038/s41419-022-05225-6

Figure Lengend Snippet: A Heatmap of significantly expressed cytokine between INRs and IRs using cytokine antibody microarray. B , C , D , and E Concentrations (pg/ml) of IL-4, IL-7, IL-15, and MCP-1 in the plasma samples obtained from INR and IR patients. F GO and KEGG analyses were conducted using DAVID for upregulated cytokine in plasma from INRs compared with IRs.

Article Snippet: The AAH-BLG-507 cytokine antibody microarray (Raybiotech, Norcross, GA, USA) was used to detect cytokines in plasma samples derived from IR and INR subjects.

Techniques: Microarray

Journal: Blood

Article Title: TCL1: a shared tumor-associated antigen for immunotherapy against B-cell lymphomas

doi: 10.1182/blood-2011-09-382838

Figure Lengend Snippet: TCL1 mRNA is hyperexpressed in multiple B-cell lymphomas. (A,C) Total RNA was extracted from normal human tissues, normal human peripheral blood B and T cells, and from primary human lymphoma cells; reverse transcribed into cDNA; and then quantitative PCR was performed for TCL1 and β-actin. (A) Expression of TCL1 mRNA relative to β-actin is shown in normal human tissues. (C) Expression of TCL1 mRNA in primary human lymphoma cells is shown relative to TCL1 expression in normal human peripheral blood B cells. Horizontal lines represent the median value for each group. P value on the top of graph represents comparison between normal donor B cells and all lymphomas. P values below the graph represent comparison between normal donor B cells and each lymphoma subtype. All P values were calculated by 2-tailed Student t test. (B,D) Expression of TCL1 mRNA in normal human tissues (B) and different human lymphoma subtypes (D) was determined from publicly available cDNA microarray datasets from the Oncomine database. The number of samples for each tissue type is shown in brackets. CLL indicates chronic lymphocytic leukemia; MCL, mantle cell lymphoma; FL, follicular lymphoma; DLBCL, diffuse large B-cell lymphoma; SMZL, splenic marginal zone B-cell lymphoma; HL, Hodgkin lymphoma; BL, Burkitt lymphoma; MM, multiple myeloma; and HCL, hairy cell leukemia.

Article Snippet: TCL1 and β-actin antibodies for Western blotting were obtained from Cell Signaling Technology.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Comparison, Microarray

Journal: Blood

Article Title: TCL1: a shared tumor-associated antigen for immunotherapy against B-cell lymphomas

doi: 10.1182/blood-2011-09-382838

Figure Lengend Snippet: TCL1 protein is hyperexpressed in multiple B-cell lymphomas. (A) Expression of TCL1 protein was determined by immunohistochemistry in formalin-fixed, paraffin-embedded primary human lymphoma tissues (FL, n = 5; CLL, n = 2; MCL, n = 3; DLBCL, n = 2; and SMZL, n = 3) and reactive human tonsils (n = 4). Images shown are the original magnification, ×100. (B) Intracellular staining for TCL1 was performed on primary human lymphoma cells and normal human peripheral blood B cells, and samples were analyzed by flow cytometry. The relative expression of TCL1 was calculated as follows: mean fluorescence intensity of TCL1 in test sample/mean fluorescence intensity of TCL1 in normal B cells. Horizontal lines represent the median value for each group. P value on the top of graph represents comparison between normal donor B cells and all lymphomas. P values below the graph represent comparison between normal donor B cells and each lymphoma subtype. All P values were calculated by 2-tailed Student t test (C) TCL1 protein expression was determined by Western blotting in 2 FL and 2 normal donor (ND) peripheral blood B cells. β-Actin protein expression was used as loading control. Vertical line inserted between lanes 2 and 3 to indicate repositioned gel lanes obtained from the same gel.

Article Snippet: TCL1 and β-actin antibodies for Western blotting were obtained from Cell Signaling Technology.

Techniques: Expressing, Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Staining, Flow Cytometry, Fluorescence, Comparison, Western Blot, Control

Journal: Blood

Article Title: TCL1: a shared tumor-associated antigen for immunotherapy against B-cell lymphomas

doi: 10.1182/blood-2011-09-382838

Figure Lengend Snippet: TCL1-specific cytotoxic T cells can be generated from normal donors. (A) PBMCs from HLA-A2+ normal donors were stimulated with overlapping 15-mer peptides spanning the entire length of the TCL1 protein (supplemental Table 1). T cells from each condition were washed and incubated with autologous CD3-depleted PBMCs as antigen-presenting cells (APC) in the presence or absence of the corresponding peptide. After 18 hours, the production of IFN-γ was determined in the supernatants by ELISA. Representative data from one of 3 normal donors tested is shown. (B) T-cell lines generated from HLA-A2+ normal donors using TCL165-79 peptide were incubated with APC in the presence or absence of the TCL165-79 peptide or control HIV Gag77-85 peptide, and intracellular cytokine assay was performed. The percentage of CD4+ and CD8+ T cells producing IFN-γ are shown. (C) TCL165-79 peptide-specific T cells were incubated with T2 cells or EBV-transformed B lymphoblastoid cell lines (IHW1003, IHW1019, IHW1089, and IHW1135) mismatched at their MHC class I locus in the presence of the TCL165-79 peptide or control HIV Gag77-85 peptide. IFN-γ production was determined as in panel A. The HLA-A alleles for each of the cell lines are shown. (D,E) A 4-hour 51Cr-release cytotoxicity assay was performed. TCL165-79 peptide-specific T cells were incubated with T2 cells alone or T2 cells pulsed with TCL165-79 peptide or control HIV Gag77-85 peptide, at various effector:target ratios. For MHC blocking assay, an effector: target ratio of 20:1 was used in the presence or absence of isotype control antibody or blocking antibodies (10 μg/mL for each antibody) against MHC class I, MHC class II, HLA-A2, or HLA-B and -C. All cytotoxicity assays were performed in triplicate wells, and mean and standard deviation are shown.

Article Snippet: TCL1 and β-actin antibodies for Western blotting were obtained from Cell Signaling Technology.

Techniques: Generated, Incubation, Enzyme-linked Immunosorbent Assay, Control, Cytokine Assay, Transformation Assay, Cytotoxicity Assay, Blocking Assay, Standard Deviation

Journal: Blood

Article Title: TCL1: a shared tumor-associated antigen for immunotherapy against B-cell lymphomas

doi: 10.1182/blood-2011-09-382838

Figure Lengend Snippet: TCL165-79 peptide-specific T cells lysed human lymphoma cell lines and primary lymphoma cells. (A,C) Intracellular staining for TCL1 was performed on tumor cell lines and normal donor peripheral blood B cells (A) and primary human lymphoma cells (C). Black histograms represent cells stained with isotype control antibody, and open histograms represent staining with TCL1 antibody. (B,D) CD8+ T cells purified from TCL165-79 peptide-specific T cell lines by magnetic cell separation were incubated with tumor cell lines and normal peripheral blood B cells (B) or primary human lymphoma cells (D). A 4-hour 51Cr-release cytotoxicity assay was performed, and the percentage of specific lysis is shown. The HLA-A2 expression for each of the cell types is shown in the figures. Data in panels B and D are representative of 3 independent experiments.

Article Snippet: TCL1 and β-actin antibodies for Western blotting were obtained from Cell Signaling Technology.

Techniques: Staining, Control, Purification, Magnetic Cell Separation, Incubation, Cytotoxicity Assay, Lysis, Expressing

Journal: Blood

Article Title: TCL1: a shared tumor-associated antigen for immunotherapy against B-cell lymphomas

doi: 10.1182/blood-2011-09-382838

Figure Lengend Snippet: TCL171-78 peptide-specific T cells can be generated from lymphoma patients. (A) PBMCs and tumor-infiltrating lymphocytes (TILs) derived from HLA-A2+ patients with CLL or FL were stimulated in vitro with TCL165-79. After 5 stimulations, IFN-γ production in response to TCL165-79 peptide was determined as described in Figure 3A. (B,E) Cytotoxic function of TCL165-79 peptide-specific T cells generated as described in panel A was tested against T2 cells pulsed with TCL165-79 peptide or HIV Gag77-85 peptide (B) or primary HLA-A2+ or HLA-A2− MCL or FL tumor cells (FL Tu) or tumor-free PBMCs from FL5 patient (FL5-PB) or HLA-A2− MCF-7 cell line (E). (C) Percentage of tetramer positive CD8+ T cells in each of the TCL1-specific T-cell lines generated from lymphoma patients is shown. (D) Expression of TCL1 in each of the primary lymphoma samples as determined by flow cytometry is shown. All cytotoxicity assays were performed in triplicate wells, and mean and standard deviation are shown. CLL1-PB-T indicates TCL1-specific CTL generated from peripheral blood T cells of CLL patient 1; FL5-PB-T, TCL1-specific CTL generated from peripheral blood T cells of FL patient 5; and FL5-TILs-T, TCL1-specific CTL generated from tumor infiltrating T cells of FL patient 5.

Article Snippet: TCL1 and β-actin antibodies for Western blotting were obtained from Cell Signaling Technology.

Techniques: Generated, Derivative Assay, In Vitro, Expressing, Flow Cytometry, Standard Deviation