Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: GADD45A suppression contributes to cardiac remodeling by promoting inflammation, fibrosis and hypertrophy

doi: 10.1007/s00018-025-05704-x

Figure Lengend Snippet: GADD45A gene expression negatively correlates with TGFB1 in human samples. ( A ) Relative quantification of the mRNA expression of GADD45A in left ventricular myocardial intraoperative biopsies obtained at surgery in patients with severe aortic stenosis (AS) compared to control subjects. The graph represents the quantification of the glyceraldehyde-3-phosphate dehydrogenase ( GAPDH )-normalized mRNA levels, expressed as a percentage of control samples. Data are presented as the mean ± SD. ( B ) Correlation coefficients between GADD45A mRNA levels and the indexed left ventricle mass (LVMI, normalized to the 2.7 power of height to neutralize differences in somatometry), mitral annular plane systolic excursion (MAPSE), and the mRNA levels of AKT3 , FABP3 , PDK4 , PDCD4 , PPARD , PTEN , and TGFB1 in the same samples. The parametric Pearson or non-parametric Spearman correlation coefficients were used to calculate the correlation for continuous variables using only complete pairs of observations. The relative transcript levels of the target genes, in arbitrary units, were used to calculate these correlation coefficients. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Small interfering RNA (siRNA)-mediated GADD45A gene silencing was carried out by transfecting AC16 cells with human GADD45A siRNA (sc-35440; Santa Cruz Biotechnology, Inc., Dallas TX, USA), using scrambled siRNA as a transfection control.

Techniques: Gene Expression, Quantitative Proteomics, Expressing, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: GADD45A suppression contributes to cardiac remodeling by promoting inflammation, fibrosis and hypertrophy

doi: 10.1007/s00018-025-05704-x

Figure Lengend Snippet: GADD45A prevents TNF-α-induced inflammation in human cardiac cells. ( A ) Relative quantification of the mRNA expression of MMP2 , MMP9 , and TGFB1 in human AC16 cardiac cells transfected with scrambled siRNA (siRNA control, siCtrl), or GADD45A siRNA (siGADD45A). ( B ) Relative quantification of the mRNA expression of CCL2 , COL1A1 , MMP2 , MMP9 , and TGFB1 in human AC16 cardiac cells transfected with LacZ-carrying or GADD45A-carrying plasmids in the presence or absence of TNF-α (TNF, 10 ng/mL, 24 h). The graphs represent the quantification of the glyceraldehyde-3-phosphate dehydrogenase ( GAPDH )-normalized mRNA levels, expressed as a percentage of control (A, siCtrl; B, LacZ) samples. Data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Small interfering RNA (siRNA)-mediated GADD45A gene silencing was carried out by transfecting AC16 cells with human GADD45A siRNA (sc-35440; Santa Cruz Biotechnology, Inc., Dallas TX, USA), using scrambled siRNA as a transfection control.

Techniques: Quantitative Proteomics, Expressing, Transfection, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: GADD45A suppression contributes to cardiac remodeling by promoting inflammation, fibrosis and hypertrophy

doi: 10.1007/s00018-025-05704-x

Figure Lengend Snippet: Deletion of Gadd45a gene in knockout mice results in hypoglycemia and reduced hepatic gluconeogenesis. Body weight ( A ), epididymal white adipose tissue (WAT) weight to body weight (BW) ratio ( B ), food intake ( C ), and plasma glucose ( D ) and triglyceride ( E ) levels in wild-type (WT) and Gadd45a knockout (KO) mice. ( F ) Pyruvate tolerance test and area under the curve (AUC). ( G ) Glucose tolerance test and area under the curve (AUC). ( H ) Relative quantification of the mRNA expression of Acox1 , Cd36 , Cpt1b , Fabp3 , Fabp4 , Fasn , Pck1 , Pdk4 , Ppara , Ppard , Ppargc1 , and Slc2a4 in the same mice. The mRNA levels were normalized to adenine phosphoribosyl transferase ( Aprt ), and are expressed as a percentage of control samples. Data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Small interfering RNA (siRNA)-mediated GADD45A gene silencing was carried out by transfecting AC16 cells with human GADD45A siRNA (sc-35440; Santa Cruz Biotechnology, Inc., Dallas TX, USA), using scrambled siRNA as a transfection control.

Techniques: Knock-Out, Clinical Proteomics, Quantitative Proteomics, Expressing, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: GADD45A suppression contributes to cardiac remodeling by promoting inflammation, fibrosis and hypertrophy

doi: 10.1007/s00018-025-05704-x

Figure Lengend Snippet: Deletion of Gadd45a gene in knockout mice boosts inflammation and fibrosis in the heart. ( A ) Relative quantification of the mRNA expression of Acta2 , Atf4 , Ccl2 , Ccn2 , Col1a1 , Il6 , Mmp2 , Mmp9 , Smad7 , Tgfb1 , and Tnf-α in wild-type (WT) and Gadd45a knockout (KO) mice. The mRNA levels were normalized to adenine phosphoribosyl transferase ( Aprt ), and are expressed as a percentage of control samples. ( B ) Western blot analysis showing the protein levels of ATF4 and phospho-SMAD3/total-SMAD3 in the same mice. The graphs represent the quantification of the protein levels normalized to vinculin, and are expressed as a percentage of control samples. Representative images of Mason’s trichrome staining ( C ) and quantification of fibrosis expressed as a percentage ( D ) in the heart of the same animals. ( E ) Western blot analysis showing the protein levels of FOS, JUN, p65, and phospho-STAT3 S727 /total-STAT3 in Gadd45a KO and WT mice, as depicted in panel B. Data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Small interfering RNA (siRNA)-mediated GADD45A gene silencing was carried out by transfecting AC16 cells with human GADD45A siRNA (sc-35440; Santa Cruz Biotechnology, Inc., Dallas TX, USA), using scrambled siRNA as a transfection control.

Techniques: Knock-Out, Quantitative Proteomics, Expressing, Control, Western Blot, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: GADD45A suppression contributes to cardiac remodeling by promoting inflammation, fibrosis and hypertrophy

doi: 10.1007/s00018-025-05704-x

Figure Lengend Snippet: The mitogen-activated protein kinases are induced in the heart of Gadd45a knockout mice. Western blot analysis showing the protein levels of phospho-ERK1/2 (extracellular signal-regulated kinase 1/2)/total-ERK1/2 ( A ), phospho-JNK (c-Jun N-terminal kinase)/total-JNK ( B ), and phospho-p38/total-p38 ( C ) mitogen-activated protein kinases (MAPK) in wild-type (WT) and Gadd45a knockout (KO) mice. The graphs represent the quantification of the protein levels normalized to vinculin, and are expressed as a percentage of control samples. Data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Small interfering RNA (siRNA)-mediated GADD45A gene silencing was carried out by transfecting AC16 cells with human GADD45A siRNA (sc-35440; Santa Cruz Biotechnology, Inc., Dallas TX, USA), using scrambled siRNA as a transfection control.

Techniques: Knock-Out, Western Blot, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: GADD45A suppression contributes to cardiac remodeling by promoting inflammation, fibrosis and hypertrophy

doi: 10.1007/s00018-025-05704-x

Figure Lengend Snippet: Deletion of Gadd45a gene in knockout mice results in substantial cardiac hypertrophy. Heart weight ( A ), and HW to body weight (BW) ( B ) and HW to tibial length (TL) ( C ) ratios in wild-type (WT) and Gadd45a knockout (KO) mice. Representative hematoxylin and eosin-stained micrographs ( D ) showing transverse sections from the left ventricle myocardium and quantification of cardiomyocyte cross-sectional areas ( E ) in the same mice. ( F ) Relative quantification of the mRNA expression of Myh6 , Myh7 , and Nppb in WT and Gadd45a KO mice. The mRNA levels were normalized to adenine phosphoribosyl transferase ( Aprt ), and are expressed as a percentage of control samples. Data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Small interfering RNA (siRNA)-mediated GADD45A gene silencing was carried out by transfecting AC16 cells with human GADD45A siRNA (sc-35440; Santa Cruz Biotechnology, Inc., Dallas TX, USA), using scrambled siRNA as a transfection control.

Techniques: Knock-Out, Staining, Quantitative Proteomics, Expressing, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: GADD45A suppression contributes to cardiac remodeling by promoting inflammation, fibrosis and hypertrophy

doi: 10.1007/s00018-025-05704-x

Figure Lengend Snippet: Cardiac apoptotic markers are induced in the heart of Gadd45a knockout mice. ( A ) Western blot analysis showing the protein levels of phospho-AKT/total-AKT, and phospho-AMPK/total-AMPK in wild-type (WT) and Gadd45a knockout (KO) mice. The graphs represent the quantification of the protein levels normalized to vinculin, and are expressed as a percentage of control samples. ( B ) Relative quantification of the mRNA expression of Bax , Bcl2 , Bcl2l1 , caspase 3 ( Casp3 ), Ddit3 , and Parp1 in WT and Gadd45a KO mice. The mRNA levels were normalized to adenine phosphoribosyl transferase ( Aprt ), and are expressed as a percentage of control samples. (C ) Western blot analysis showing the protein levels of BAX, BCL2, cleaved caspase 3, CHOP and cleaved PARP1 in the same mice, as depicted in panel A. Data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Small interfering RNA (siRNA)-mediated GADD45A gene silencing was carried out by transfecting AC16 cells with human GADD45A siRNA (sc-35440; Santa Cruz Biotechnology, Inc., Dallas TX, USA), using scrambled siRNA as a transfection control.

Techniques: Knock-Out, Western Blot, Control, Quantitative Proteomics, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: GADD45A suppression contributes to cardiac remodeling by promoting inflammation, fibrosis and hypertrophy

doi: 10.1007/s00018-025-05704-x

Figure Lengend Snippet: Echocardiographic data of wild-type (WT) or Gadd45a knockout (KO) male mice

Article Snippet: Small interfering RNA (siRNA)-mediated GADD45A gene silencing was carried out by transfecting AC16 cells with human GADD45A siRNA (sc-35440; Santa Cruz Biotechnology, Inc., Dallas TX, USA), using scrambled siRNA as a transfection control.

Techniques: Knock-Out

Journal: Nature Communications

Article Title: FTO deficiency in older livers exacerbates ferroptosis during ischaemia/reperfusion injury by upregulating ACSL4 and TFRC

doi: 10.1038/s41467-024-49202-3

Figure Lengend Snippet: a Pie chart showing the distribution and relative proportion of differential peaks between the control and FTO groups according to m6A RNA sequencing. b Pie chart showing the percentages of up/downregulated differential peaks. c Representative FTO potential binding motifs. d Bubble chart showing the enriched signaling pathways with down-regulated m6A peaks. Binomial distribution test (two-sided), P < 0.05 was considered statistically significant. e Integrated genome browser views show the m6A modifications of Acsl4 and Tfrc in the control and FTO groups. f MeRIP-qPCR quantitative analysis of the fold enrichment of Acsl4 and Tfrc m6A levels by immunoprecipitation with a specific m6A antibody in senescent THLE2 cells transfected with control and FTO (two-tailed t -test). g Integrated genome browser views show the binding peaks by FTO in Acsl4 and Tfrc via FTO-CLIP-seq. h RT‒qPCR validation of target gene expression after overexpression or silencing of FTO in primary hepatocytes (two-tailed t -test ). i Effects of FTO on the expression of ACSL4 and TFRC in primary hepatocytes via western blotting, three independent cell experiments. j Representative images of IHC staining (magnification, × 200) for FTO, ACSL4 and TFRC in clinical liver specimens, three independent clinical human samples per group. k, l Effects of FTO on the half-life of Acsl4 and Tfrc mRNA in senescent THLE2 cells via RT‒qPCR. m Effects of FTO on the regulation of YTHDC2 on ACSL4 via western blotting. n Effects of FTO on the regulation of YTHDF2 on TFRC via western blotting. m, n Three independent cell experiments. o Effects of FTO on the enrichment of Acsl4 mRNA by YTHDC2 via RIP assays, one-way ANOVA followed by multiple comparisons. p Effects of FTO on the enrichment of Tfrc mRNA by YTHDF2 via RIP assays, one-way ANOVA followed by multiple comparisons. Statistic data are presented as the mean±SD, error bars represent the means of three independent experiments. P < 0.05 was considered statistically significant, source data are provided as a Source Data file.

Article Snippet: Add YTHDF2 or YTHDC2 antibody (CST, Danvers, MA, USA) to pre-washed Protein A/G magnetic beads (Thermo Fisher, USA) and incubate with rotation for 2 h at room temperature.

Techniques: Control, RNA Sequencing, Binding Assay, Protein-Protein interactions, Immunoprecipitation, Transfection, Two Tailed Test, Biomarker Discovery, Targeted Gene Expression, Over Expression, Expressing, Western Blot, Immunohistochemistry

Journal: Nature Communications

Article Title: FTO deficiency in older livers exacerbates ferroptosis during ischaemia/reperfusion injury by upregulating ACSL4 and TFRC

doi: 10.1038/s41467-024-49202-3

Figure Lengend Snippet: a Pie chart showing the distribution and relative proportion of differential peaks between the control and FTO groups according to m6A RNA sequencing. b Pie chart showing the percentages of up/downregulated differential peaks. c Representative FTO potential binding motifs. d Bubble chart showing the enriched signaling pathways with down-regulated m6A peaks. Binomial distribution test (two-sided), P < 0.05 was considered statistically significant. e Integrated genome browser views show the m6A modifications of Acsl4 and Tfrc in the control and FTO groups. f MeRIP-qPCR quantitative analysis of the fold enrichment of Acsl4 and Tfrc m6A levels by immunoprecipitation with a specific m6A antibody in senescent THLE2 cells transfected with control and FTO (two-tailed t -test). g Integrated genome browser views show the binding peaks by FTO in Acsl4 and Tfrc via FTO-CLIP-seq. h RT‒qPCR validation of target gene expression after overexpression or silencing of FTO in primary hepatocytes (two-tailed t -test ). i Effects of FTO on the expression of ACSL4 and TFRC in primary hepatocytes via western blotting, three independent cell experiments. j Representative images of IHC staining (magnification, × 200) for FTO, ACSL4 and TFRC in clinical liver specimens, three independent clinical human samples per group. k, l Effects of FTO on the half-life of Acsl4 and Tfrc mRNA in senescent THLE2 cells via RT‒qPCR. m Effects of FTO on the regulation of YTHDC2 on ACSL4 via western blotting. n Effects of FTO on the regulation of YTHDF2 on TFRC via western blotting. m, n Three independent cell experiments. o Effects of FTO on the enrichment of Acsl4 mRNA by YTHDC2 via RIP assays, one-way ANOVA followed by multiple comparisons. p Effects of FTO on the enrichment of Tfrc mRNA by YTHDF2 via RIP assays, one-way ANOVA followed by multiple comparisons. Statistic data are presented as the mean±SD, error bars represent the means of three independent experiments. P < 0.05 was considered statistically significant, source data are provided as a Source Data file.

Article Snippet: Add YTHDF2 or YTHDC2 antibody (CST, Danvers, MA, USA) to pre-washed Protein A/G magnetic beads (Thermo Fisher, USA) and incubate with rotation for 2 h at room temperature.

Techniques: Control, RNA Sequencing, Binding Assay, Protein-Protein interactions, Immunoprecipitation, Transfection, Two Tailed Test, Biomarker Discovery, Targeted Gene Expression, Over Expression, Expressing, Western Blot, Immunohistochemistry