Journal: International journal of oral science

Article Title: WNT7A promotes tumorigenesis of head and neck squamous cell carcinoma via activating FZD7/JAK1/STAT3 signaling.

doi: 10.1038/s41368-024-00279-y

Figure Lengend Snippet: Fig. 4 WNT7A upregulated expression of STAT3 target genes in HNSCC cells. a Heatmap of differentially expressed genes based on RNA- seq data. The top 30 deregulated genes were selected for further analysis. b, c Validation of the promote proliferation, self-renewal, and anti- apoptosis correlated genes by real-time RT-PCR. d Design of ChIP-qPCR primers by using Cistrome Data Browser. e ChIP-qPCR analysis was performed to examine the enrichment of STAT4, HCAR2, and BIRC3 after overexpression of WNT7A. The immunoprecipitation of the STAT3 was compared to that of the IgG antibody and IRF1 was a positive control in the analysis. Data shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.000 1

Article Snippet: . . Huang et al. 9 International Journal of Oral Science (2024) 16:7 Chromatin immunoprecipitation (ChIP-qPCR) ChIP assays were conducted using an anti-STAT3 antibody (Table S4 shows the antibodies used for ChIP) and Protein G Magnetic Beads (HY-K0204, MCE, USA) to pull down DNA fragments bound to STAT3.

Techniques: Expressing, RNA Sequencing, Biomarker Discovery, Quantitative RT-PCR, ChIP-qPCR, Over Expression, Immunoprecipitation, Positive Control

Journal: International journal of oral science

Article Title: WNT7A promotes tumorigenesis of head and neck squamous cell carcinoma via activating FZD7/JAK1/STAT3 signaling.

doi: 10.1038/s41368-024-00279-y

Figure Lengend Snippet: Fig. 6 WNT7A may activate STAT3 signaling pathway through FZD7/JAK1. a Western blot analysis demonstrated that overexpression of WNT7A in HN30 and HN6 cells leaded to increased Tyr1034/1035 phosphorylation of JAK1, while total protein levels of JAK1 remain unchanged. b Protein network prediction using the STRING database identified FZD5 and FZD7 as potential receptor candidates for WNT7A. c Analysis of single-cell sequencing data (GSE103322) revealed higher enrichment of FZD7 in cancer cells. d, e Co-immunoprecipitation results confirm interactions between WNT7A, FZD7, and JAK1. f Co-localization assay by staining FZD7 (red), JAK1 (green), and nucleus (DAPI)

Article Snippet: . . Huang et al. 9 International Journal of Oral Science (2024) 16:7 Chromatin immunoprecipitation (ChIP-qPCR) ChIP assays were conducted using an anti-STAT3 antibody (Table S4 shows the antibodies used for ChIP) and Protein G Magnetic Beads (HY-K0204, MCE, USA) to pull down DNA fragments bound to STAT3.

Techniques: Western Blot, Over Expression, Phospho-proteomics, Single Cell, Sequencing, Immunoprecipitation, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Downregulation of PDIA3 inhibits gastric cancer cell growth through cell cycle regulation.

doi: 10.1016/j.biopha.2024.116336

Figure Lengend Snippet: Fig. 8. STAT3 was associated with PDIA3-mediated cyclin G1 regulation. (A) The protein expressions of STAT3, phosphor-STAT3 (Tyr705) and cyclin G1 after downregulation of PDIA3 by sgRNA-2. (B) The expression of cyclin G1 after treatment of a STAT3 inhibitor cryptotanshinone in TMK1 and AGS. (C) The expression of cyclin G1 after siSTAT3 in TMK1 and AGS.

Article Snippet: The inhibitor for STAT3, cryptotanshinone, was purchased from MedChemExpress LLC. (Shanghai, China).

Techniques: Expressing

Journal: Cell Death & Disease

Article Title: Hippo kinases Mst1 and Mst2 maintain NK cell homeostasis by orchestrating metabolic state and transcriptional activity

doi: 10.1038/s41419-024-06828-x

Figure Lengend Snippet: A , B Flow cytometry analysis (left) and MFI intensity (right) demonstrate the expression levels of p-STAT5, p-S6, p-AKT473 and p-Erk in NK cells from Mst1/2 fl/fl and Mst1/2 fl/fl CD122 Cre mice upon stimulation with or without IL-15 (50 ng/mL) for 30 min ( n ≥ 4). C Gene-set enrichment analysis of RNA-seq data shows the enrichment of hallmark-IL6-JAK-STAT3-signaling related genes in Mst1/2-deficient NK cells compared to WT NK cells. D Flow cytometry analysis (left) and MFI intensity (right) reveals the expression level of p-STAT3 (Ser727) in NK cells from Mst1/2 fl/fl and Mst1/2 fl/fl CD122 Cre mice upon stimulation with or without IL-15 (50 ng/mL) for 30 min ( n = 4). E ChIP-qPCR analysis demonstrated binding of p-STAT3 conserved motifs in the Tcf7 5′ regulatory region (−17.5 kb), while no binding was observed in a region without p-STAT3-binding motifs (+0.2 kb), serving as a negative control. The experiments were using sorted NK cells from WT mice with anti-p-STAT3 antibody or isotype-matched IgG. F – I WT splenocytes were treated with DMSO, Mst1 inhibitor XMU-MP-1 (1 mM, MedChemExpress) or p-STAT3 inhibitor Stattic (1 mM, MedChemExpress) for 24 h, followed by flow cytometry analysis. The representative plots (left) and summary data (right) show the expression level of p-STAT3 (F), TCF1 ( G ), total cellular ROS ( H ) and the percentage of 7AAD + NK cells ( I ) ( n = 4). J Schematic illustration of Mst1/2 regulate NK cell survival and function via controlling metabolic state and transcriptional activity. Data A , B and D – I are representative of three independent experiments with similar results.

Article Snippet: F – I WT splenocytes were treated with DMSO, Mst1 inhibitor XMU-MP-1 (1 mM, MedChemExpress) or p-STAT3 inhibitor Stattic (1 mM, MedChemExpress) for 24 h, followed by flow cytometry analysis.

Techniques: Flow Cytometry, Expressing, RNA Sequencing, ChIP-qPCR, Binding Assay, Negative Control, Activity Assay

Journal: Cells, tissues, organs

Article Title: The role of epithelial Stat3 in amelogenesis during mouse incisor renewal

doi: 10.1159/000486745

Figure Lengend Snippet: (A) Illustration of the adult mouse hemi-mandible showing the incisor and molars, as well as the mineralized dentin and enamel in the incisor. (A’) The proximal region of the incisor denoting the labial and lingual cervical loop (laCL and liCL, respectively, highlighted by dashed, red lines). (A’’) Magnified illustration of the laCL indicating the outer enamel epithelium (OEE), inner enamel epithelium (IEE), transit amplifying (TA) region, and stellate reticulum (SR). (B–I) Immunofluorescence staining for STAT3 in wildtype mouse teeth (sagittal views) at E14.5, E16.5, P1, and 6 weeks showed the presence of STAT3 in developing incisors and molars, and adult incisors. The epithelial component (i.e., cells that ultimately generate enamel) of developing teeth are outlined (dotted red line) with the exception of 6-week old molar where the entire tooth is outlined (I). (J,K’) H&E histological staining of control (Stat3fl/fl) and mutant (Krt14Cre;Stat3fl/fl) adult mice mandibular incisors showed little or no enamel matrix in mutants compared to controls.

Article Snippet: Primary antibodies used were as follows: anti-rabbit STAT3 (1:200; Cat#7197, ProSci, used to detect STAT3 expression in wildtype specimens), anti-rabbit STAT3 (Phospho-Stat3 (Tyr705) (D3A7), Cat#9145S, Cell signaling, used to detect STAT3 expression in Stat3 fl/fl and K rt14 Cre/+ ; Stat3 fl/fl mice), anti-rabbit AMELX (amelogenin; 1:200; Cat#ab54507, Abcam), anti-rabbit AMBN (ameloblastin; 1:200; Cat#ab116347, Abcam), anti-kallikrein-4 or KLK4 (1:200; Abcam, Cat#ab71234, Abcam), enamelin or ENAM (1:200; Cat#sc-33107, Santa Cruz), and KI67 (1:200, Cat#RM-9106, Thermo Scientific).

Techniques: Immunofluorescence, Staining, Control, Mutagenesis