sox4 specific antibody thermo pa5-72852  (Thermo Fisher)

Journal: Journal of Translational Medicine

Article Title: ELF3-regulated PES1 targets VEGFR2 to mediate angiogenesis and retinal inner barrier injury in diabetic retinopathy

doi: 10.1186/s12967-025-07334-0

Figure Lengend Snippet: ELF3 is a newly discovered transcription factor that regulates PES1. ( A ) The UCSC database predicts the transcription factors that may regulate PES1. ( B ) The expression of ELF3 and SOX4 in HRMECs. Student t-test. n = 3/group. * p < 0.05 versus NG. ( C , D ) The expression of ELF3 and SOX4 in HRMECs. One-way ANOVA. n = 3/group. * p < 0.05 versus HG + NC siRNA. ( E , F ) The mRNA level of ELF3 and SOX4 in HRMECs. One-way ANOVA. n = 3/group. * p < 0.05 versus HG + NC siRNA. ( G ) Enrichment ability of ELF3 in PES1 promoter regions. Student t-test. n = 3/group. * p < 0.05 versus IgG. ( H ) Luciferase activity of PES1 promoter transfected with ELF3 overexpress plasmids or control plasmids. One-way ANOVA. n = 3/group. * p < 0.05 versus ELF3 NC group

Article Snippet: Primary or secondary antibodies were summarized as follows: PES1(1:5000, 13553-1-AP; Proteintech, Wuhan, China; 1:100, sc-166300; Santa Cruz); VEGFA (1:1000, 19003-1-AP; Proteintech); NOX4 (1:5000, 14347-1-AP; Proteintech); VE-cadherin (1:1000, 66804-1-AP; Proteintech; 1:2000, YM8373; Immunoway, China); Occludin (1:5000, 27260-1-AP; Proteintech); SOD1 (1:10000, 10269-1-AP; Proteintech); VEGFR2 (1:1000, 26415-1-AP; Proteintech; 1:1000, R380984; Zenbio, China); ELF3 (1:1000, R389278; Zenbio); SOX4 (1:1000, CSB-PA022431LA01HU; CUSABIO, China); β-actin (1:3000, GB15003-100; Servicebio, China).

Techniques: Expressing, Luciferase, Activity Assay, Transfection, Control

Journal: Frontiers in Immunology

Article Title: The cancer-associated fibroblasts interact with malignant T cells in mycosis fungoides and promote the disease progression

doi: 10.3389/fimmu.2024.1474564

Figure Lengend Snippet: The interaction between malignant T cell and CAFs with SOX4-related mechanisms in the process. (A) Malignant T cells in MF induce the activation and formation of CAFs, which, in turn, modulating cancer progression. (B) Activation of SOX4 expression in malignant T cells. IL-6 generated by CAFs, activating the JAK2/STAT3 pathway in malignant T cells, positively regulated the transcription of SOX4, further activating the PI3K/Akt, Wnt/β-catenin, Notch, and Hedgehog signaling pathway, hence promoting tumorigenesis. (C) Activation of SOX4 expression in CAFs. TGF-β generated by malignant cells activates the TGF-β signaling in NFs or epithelial cells, leading to the phosphorylation of SMAD2/3, which promotes SOX4 expression after translocating to the nucleus. By interacting with β-catenin and ERG, SOX4 induces the expression of EZH2, which might directly regulate TWIST1 and modulating the EMT procession, reprograming other cells into CAFs. The SOX4–EZH2 axis also modifies the promoter region of tumor-suppressor microRNAs (miRNAs), including miR-31, miR-212/132, and miR-129-2. By repressing the transcription of these miRNAs, SOX4 involved in the inhibition of cell proliferation and migration.

Article Snippet: Primary antibodies used in immunohistochemistry (IHC) and immunofluorescence (IF) are listed as follows: cluster of differentiation 4 (CD4) (1:100, Abcam, ab133616, clone EPR6855), CD8 (1:1,000, Proteintech, 66868-1-lg, clone 1G2B10), SRY-related high-mobility-group box 4 (SOX4) (1:200, CUSABIO, CSBPA022431LA01HU), Nuclear Receptor Subfamily 4 Group A Member 1 (NR4A1) (1:200, Proteintech, 25851-1-AP), CD103 (1:100, Abcam, ab224202, EPR22590-27), Fibroblast Activation Protein Alpha (FAP) (1:250, Abcam, ab207178, EPR20021), Twist Family BHLH Transcription Factor 1 (TWIST1) (1:200, Proteintech, 25465-1-AP), Matrix Metallopeptidase 2 (MMP2) (1:200, Proteintech, 10373-2-AP), and Interleukin 6 (IL-6) (1:200, Proteintech, 21865-1-AP).

Techniques: Activation Assay, Expressing, Generated, Phospho-proteomics, Inhibition, Migration

Journal: Frontiers in Immunology

Article Title: The cancer-associated fibroblasts interact with malignant T cells in mycosis fungoides and promote the disease progression

doi: 10.3389/fimmu.2024.1474564

Figure Lengend Snippet: Transcriptional profiles of fibroblast subpopulations in patients with MF and HC. (A) UMAP visualization of seven fibroblast subpopulations. (B) Heatmap for gene expression levels of top 10 SDE genes of fibroblast subtypes. (C) UMAP plots color-coded for the expression of marker genes for CAF subtypes. (D) Density plot showing the distribution of each fibroblast subpopulation along the pseudotime inferred by analysis with Monocle2. (E) NFs were selected as the start cells, color key from dark to bright indicates cancer progression from the early to the late stage. (F) Heatmap showing the dynamic changes in gene expression along the pseudotime of fibroblasts. Color key from blue to red indicates relative expression levels from low to high. (G) Immunohistochemical stain of SOX4, FAP, IL-6, TWIST1, and MMP2 in skin biopsies of patch stage MF lesion, tumor stage MF lesion, and LP lesion. Original magnification was ×20. Scale bar, 100 μm. (H) Representative IF staining of FAP (red), SOX4 (green), and IL-6 (magenta) on paraffin-embedded tissue samples of tumor stage lesion from MF2 patient. DAPI (blue) was used to visualize cell nuclei. Scale bar, 50 μm. (I) GSVA analysis indicates enriched pathways of each subset of CD4+ T cell.

Article Snippet: Primary antibodies used in immunohistochemistry (IHC) and immunofluorescence (IF) are listed as follows: cluster of differentiation 4 (CD4) (1:100, Abcam, ab133616, clone EPR6855), CD8 (1:1,000, Proteintech, 66868-1-lg, clone 1G2B10), SRY-related high-mobility-group box 4 (SOX4) (1:200, CUSABIO, CSBPA022431LA01HU), Nuclear Receptor Subfamily 4 Group A Member 1 (NR4A1) (1:200, Proteintech, 25851-1-AP), CD103 (1:100, Abcam, ab224202, EPR22590-27), Fibroblast Activation Protein Alpha (FAP) (1:250, Abcam, ab207178, EPR20021), Twist Family BHLH Transcription Factor 1 (TWIST1) (1:200, Proteintech, 25465-1-AP), Matrix Metallopeptidase 2 (MMP2) (1:200, Proteintech, 10373-2-AP), and Interleukin 6 (IL-6) (1:200, Proteintech, 21865-1-AP).

Techniques: Gene Expression, Expressing, Marker, Immunohistochemical staining, Staining

Journal: Frontiers in Immunology

Article Title: The cancer-associated fibroblasts interact with malignant T cells in mycosis fungoides and promote the disease progression

doi: 10.3389/fimmu.2024.1474564

Figure Lengend Snippet: Cell–cell communications between T cells and fibroblasts. (A) Expression levels of SOX4 and IL-6R for T-cell subtypes are plotted onto the UMAP plots. (B) Violin plots show the expression of specific markers in T-cell subtypes (C01–C06, C08, C10, C11, C13, and C16). (C) Overall survival (OS) and progression-free survival (PFS) of patients with MF stratified by SOX4 expression from a publicly available GEO MF cohort of 49 RNA-seq samples via Kaplan–Meier survival analysis. (D, E) The interaction number of T cells and fibroblast subpopulations in plaque and tumor stage lesion of MF1 patient. The thickness of the line represents the interaction number between the subpopulations estimated by CellPhoneDB. (F) Summary of selected immune-associated ligand–receptor pairs between malignant T cells and CAFs in all samples using CellPhoneDB. The size of each dot denotes the p-value. The color gradient denotes the degree of interaction. (G) The major interactions between malignant T cells (C01 and C11) and CAF subtypes.

Article Snippet: Primary antibodies used in immunohistochemistry (IHC) and immunofluorescence (IF) are listed as follows: cluster of differentiation 4 (CD4) (1:100, Abcam, ab133616, clone EPR6855), CD8 (1:1,000, Proteintech, 66868-1-lg, clone 1G2B10), SRY-related high-mobility-group box 4 (SOX4) (1:200, CUSABIO, CSBPA022431LA01HU), Nuclear Receptor Subfamily 4 Group A Member 1 (NR4A1) (1:200, Proteintech, 25851-1-AP), CD103 (1:100, Abcam, ab224202, EPR22590-27), Fibroblast Activation Protein Alpha (FAP) (1:250, Abcam, ab207178, EPR20021), Twist Family BHLH Transcription Factor 1 (TWIST1) (1:200, Proteintech, 25465-1-AP), Matrix Metallopeptidase 2 (MMP2) (1:200, Proteintech, 10373-2-AP), and Interleukin 6 (IL-6) (1:200, Proteintech, 21865-1-AP).

Techniques: Expressing, RNA Sequencing