p hsf1 ser326  (Bioss)

Journal: Molecular Therapy. Nucleic Acids

Article Title: Aberrant HSF1 signaling activation underlies metformin amelioration of myocardial infarction in mice

doi: 10.1016/j.omtn.2022.07.009

Figure Lengend Snippet: HSF1 had protective effect on mouse myocardium (A) Expression levels of p-HSF1Ser326/HSF1 in tissues adjacent to infarcted myocardium in different treatment groups. (B) mRNA levels of HSF1 in tissues near infarcted myocardium in different treatment groups. (n = 3; SD) (C) Subcellular distribution of HSF1 in mouse cardiomyocytes. Red represents HSF1 fluorescence and blue represents DAPI. Scale bar, 50 μm. (D) Expression levels of HSF1 in the cytoplasm and nucleus, respectively. Cytoplasmic proteins were homogenized with β-actin as an internal reference, and nuclear proteins were homogenized with histone H3 as an internal reference. (E) Masson staining of representative left ventricular section. The right panel shows infarct sizes. LV, left ventricle; INF, infarct area. Scale bar, 2,000 μm; n = 3; ∗∗∗p < 0.001. (F) Representative pictures of transmission electron microscopy of tissue adjacent to infarcted myocardium in wild-type mice (WT) and HSF1-knockout homozygous mice (HSF1−/−). Scale bar, 1 μm.

Article Snippet: The primary antibodies used were anti-HSF1 and p-HSF1 Ser326 (Cell Signaling Technology, USA), anti-β-actin (Cell Signaling Technology, USA), anti-LC3 (Cell Signaling Technology, USA), anti-p62 (Cell Signaling Technology, USA), Histone H3 (Cell Signaling Technology, USA), anti-mTOR and anti-p-mTOR Ser2448 (Abcam, USA), anti-AMPK and anti-p-AMPK Thr172 (Abcam, USA).

Techniques: Expressing, Fluorescence, Staining, Transmission Assay, Electron Microscopy, Knock-Out

Journal: Molecular Therapy. Nucleic Acids

Article Title: Aberrant HSF1 signaling activation underlies metformin amelioration of myocardial infarction in mice

doi: 10.1016/j.omtn.2022.07.009

Figure Lengend Snippet: HSF1 was the upstream regulator of AMPK/mTOR pathway (A) The mRNA expression levels of HSF1, AMPK, and mTOR in the cardiomyocytes of HSF1 overexpression group and its control group (Ad-HSF1 and Ad-NC), HSF1 knockdown group and its control group (si-HSF1 and si-NC) mice. (n = 3; SD; ∗∗∗p < 0.001) (B) Scatterplot of the correlation analysis between HSF1 and AMPK mRNA expression in mouse cardiomyocytes, measured by qPCR. (C) Punctate accumulation of GFP-LC3 in mouse cardiomyocytes at 48 h in different treatment groups (Ad-NC, Ad-HSF1, si-NC, and si-HSF1). (D) Expression levels of HSF1, AMPK/mTOR pathway, autophagy-related protein p62, and LC3II/I in mouse cardiomyocytes. (E) Expression levels of HSF1 and AMPK proteins in mouse cardiomyocytes after overexpression and knockdown of AMPK (Ad-NC, Ad-AMPK, si-NC, and si-AMPK). (F) Expression levels of mTOR and autophagy-related proteins p62, LC3II/I in mouse cardiomyocytes after overexpression and knockdown of AMPK or HSF1 (Ad-HSF1, si-HSF1, Ad-AMPK, and si-AMPK).

Article Snippet: The primary antibodies used were anti-HSF1 and p-HSF1 Ser326 (Cell Signaling Technology, USA), anti-β-actin (Cell Signaling Technology, USA), anti-LC3 (Cell Signaling Technology, USA), anti-p62 (Cell Signaling Technology, USA), Histone H3 (Cell Signaling Technology, USA), anti-mTOR and anti-p-mTOR Ser2448 (Abcam, USA), anti-AMPK and anti-p-AMPK Thr172 (Abcam, USA).

Techniques: Expressing, Over Expression, Control, Knockdown

Journal: Molecular Therapy. Nucleic Acids

Article Title: Aberrant HSF1 signaling activation underlies metformin amelioration of myocardial infarction in mice

doi: 10.1016/j.omtn.2022.07.009

Figure Lengend Snippet: AMPKα2 promoter reporters were transactivated by HSF1 (A) Diagram shows the relative positions of full-length (FL) and fragments of AMPKα2 promoter reporters. (B and C) Reporter assay response of AMPKα2 promoter FL and the individual fragments was investigated. (D) Reporter assays of the P2 fragment of the AMPKα2 promoter containing a mutated TFBM as indicated. (E) A schematic illustrates the relative positions of qPCR probes to putative motif (pm) for ChIP-QPCR experiments. (F) Antibody-pulled down chromatins were analyzed by qPCR. RLU, relative light unit. Data were mean ± SD of three independent experiments. (n = 3; SD; ∗∗∗p < 0.001)

Article Snippet: The primary antibodies used were anti-HSF1 and p-HSF1 Ser326 (Cell Signaling Technology, USA), anti-β-actin (Cell Signaling Technology, USA), anti-LC3 (Cell Signaling Technology, USA), anti-p62 (Cell Signaling Technology, USA), Histone H3 (Cell Signaling Technology, USA), anti-mTOR and anti-p-mTOR Ser2448 (Abcam, USA), anti-AMPK and anti-p-AMPK Thr172 (Abcam, USA).

Techniques: Reporter Assay, ChIP-qPCR

Journal: Molecular Therapy. Nucleic Acids

Article Title: Aberrant HSF1 signaling activation underlies metformin amelioration of myocardial infarction in mice

doi: 10.1016/j.omtn.2022.07.009

Figure Lengend Snippet: The combination of metformin-HSF1 showed a stronger binding effect than the combination of metformin-AMPK (A and B) Schematic diagram of metformin ligand binding AMPK (A) and HSF1 (B) protein (pymol mapping); the protein is displayed in cartoon form. (C and D) Schematic diagram of the amino acid sites of metformin ligand binding to AMPK (C) and HSF1 (D) protein (pymol mapping). (E and F) Concentration gradient curve of compound metformin and protein AMPK (E) and HSF1 (F). (G) Binding curves of two proteins (AMPK and HSF1) at 3,200 nM and metformin, respectively. (H) Histogram of the maximum binding signal between the two proteins at 3,200 nM and metformin on the chip.

Article Snippet: The primary antibodies used were anti-HSF1 and p-HSF1 Ser326 (Cell Signaling Technology, USA), anti-β-actin (Cell Signaling Technology, USA), anti-LC3 (Cell Signaling Technology, USA), anti-p62 (Cell Signaling Technology, USA), Histone H3 (Cell Signaling Technology, USA), anti-mTOR and anti-p-mTOR Ser2448 (Abcam, USA), anti-AMPK and anti-p-AMPK Thr172 (Abcam, USA).

Techniques: Binding Assay, Ligand Binding Assay, Concentration Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: Aberrant HSF1 signaling activation underlies metformin amelioration of myocardial infarction in mice

doi: 10.1016/j.omtn.2022.07.009

Figure Lengend Snippet: Comparison of the docking energy of metformin with AMPK and HSF1

Article Snippet: The primary antibodies used were anti-HSF1 and p-HSF1 Ser326 (Cell Signaling Technology, USA), anti-β-actin (Cell Signaling Technology, USA), anti-LC3 (Cell Signaling Technology, USA), anti-p62 (Cell Signaling Technology, USA), Histone H3 (Cell Signaling Technology, USA), anti-mTOR and anti-p-mTOR Ser2448 (Abcam, USA), anti-AMPK and anti-p-AMPK Thr172 (Abcam, USA).

Techniques: Comparison

Journal: Molecular Therapy. Nucleic Acids

Article Title: Aberrant HSF1 signaling activation underlies metformin amelioration of myocardial infarction in mice

doi: 10.1016/j.omtn.2022.07.009

Figure Lengend Snippet: Analyze the affinity data of metformin with two proteins by SPR

Article Snippet: The primary antibodies used were anti-HSF1 and p-HSF1 Ser326 (Cell Signaling Technology, USA), anti-β-actin (Cell Signaling Technology, USA), anti-LC3 (Cell Signaling Technology, USA), anti-p62 (Cell Signaling Technology, USA), Histone H3 (Cell Signaling Technology, USA), anti-mTOR and anti-p-mTOR Ser2448 (Abcam, USA), anti-AMPK and anti-p-AMPK Thr172 (Abcam, USA).

Techniques:

Journal: Molecular Therapy. Nucleic Acids

Article Title: Aberrant HSF1 signaling activation underlies metformin amelioration of myocardial infarction in mice

doi: 10.1016/j.omtn.2022.07.009

Figure Lengend Snippet: Mechanistic diagram of metformin (via HSF1 and AMPK) protecting mouse myocardium

Article Snippet: The primary antibodies used were anti-HSF1 and p-HSF1 Ser326 (Cell Signaling Technology, USA), anti-β-actin (Cell Signaling Technology, USA), anti-LC3 (Cell Signaling Technology, USA), anti-p62 (Cell Signaling Technology, USA), Histone H3 (Cell Signaling Technology, USA), anti-mTOR and anti-p-mTOR Ser2448 (Abcam, USA), anti-AMPK and anti-p-AMPK Thr172 (Abcam, USA).

Techniques: