Journal: Molecular Metabolism

Article Title: ANKRD53 is downregulated in human obesity and coordinates lipolysis with mitochondrial oxidative metabolism in adipocytes

doi: 10.1016/j.molmet.2026.102330

Figure Lengend Snippet: ANKRD53 expression in human adipose tissue inversely associates with obesity and adverse clinical metabolic traits. (a) Relative ANKRD53 expression in subcutaneous (SAT) and visceral (VAT) adipose tissues from non-obesity (BMI <30 kg/m 2 ; n = 87) and obesity (BMI ≥30 kg/m 2 ; n = 149) individuals. (b, c) Spearman correlations of ANKRD53 expression in SAT (b) and VAT (c) with BMI, fat mass, waist-to-hip ratio (WHR) and subcutaneous fat area (SFA) or visceral fat area (VFA). (d, e) Spearman correlations of ANKRD53 expression in SAT (d) and VAT (e) with fasting plasma glucose (FPG), HbA1c, fasting insulin (FINS), and HOMA-IR (homeostasis model assessment of insulin resistance). (f, g) Spearman correlations of ANKRD53 expression in SAT (f) and VAT (g) with triglycerides (TG), HDL-cholesterol (HDL-C), and serum adiponectin. (h) Regional association plots at the ANKRD53 locus (chromosome 2), displaying single-nucleotide polymorphisms (SNPs) associated with BMI (top) and triglyceride levels (bottom). Data were obtained from the Common Metabolic Disease Knowledge Portal ( https://t2d.hugeamp.org/ ). (i) KEGG pathway enrichment analysis of genes positively correlated with ANKRD53 expression in SAT (669 genes; r > 0.4, p < 0.05). (j) KEGG pathway enrichment analysis of genes positively correlated with ANKRD53 expression in VAT (840 genes; r > 0.4, p < 0.05). Gene expression was quantified from RNA-seq data (n = 236 individuals) and normalized as fragments per kilobase of transcript per million mapped reads (FPKM). Spearman correlation coefficients (r) Spearman correlation coefficients (r), corresponding p -values and FDR values are shown. Statistical analysis was performed using a two-tailed unpaired Student's t-test. Data are presented as mean ± SEM. FDR values were adjusted using the Benjamini-Hochberg method. ∗∗ p < 0.01. ∗∗∗ p < 0.001. FDR <0.05 is considered statistically significant.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST and incubated overnight at 4 °C with the following primary antibodies: ANKRD53 (Proteintech, 24283-1-AP), Tubulin (Sigma–Aldrich, T6199), phospho-HSL (Ser660) (Cell Signaling Technology, 45804), HSL (Cell Signaling Technology, 4107), Phospho-PKA Substrate (Cell Signaling Technology, 9624), and Histone H3 (Cell Signaling Technology, 4499).

Techniques: Expressing, Clinical Proteomics, Gene Expression, RNA Sequencing, Two Tailed Test

Journal: Molecular Metabolism

Article Title: ANKRD53 is downregulated in human obesity and coordinates lipolysis with mitochondrial oxidative metabolism in adipocytes

doi: 10.1016/j.molmet.2026.102330

Figure Lengend Snippet: Selective expression and regulation of ANKRD53 in human primary adipocytes by metabolic cues (a) Relative Ankrd5 3 mRNA expression in mouse adipose tissue, hypothalamus and testis (n = 6). (b) Relative ANKRD5 3 mRNA expression across human adipose depots, including superficial neck (n = 14), deep neck (n = 12), SAT (n = 12) and VAT (n = 12). (c) Relative ANKRD5 3 mRNA expression in paired mature adipocytes (MF) and stromal vascular fractions (SVFs) isolated from human SAT (n = 3). (d) Relative ANKRD5 3 mRNA expression during adipogenic differentiation of human SVFs in vitro (n = 4). (e) Dot plot of ANKRD53 expression across major cell populations in human white adipose tissue. Dot size represents the proportion of cells expressing ANKRD53, and color intensity reflects scaled average expression. (f) Relative ANKRD5 3 mRNA expression in mature human primary adipocytes treated with forskolin (10 μM, 6 h) (n = 4). (g) Relative ANKRD5 3 mRNA expression in mature human primary adipocytes treated with rosiglitazone (1 μM) (n = 6). (h) Relative ANKRD53 expression in SAT from healthy individuals under 2h (fed) or 24h (fast) after a standard meal (postprandial; GSE154612 ; n = 11). (i) Relative ANKRD53 expression in SAT from obese individuals before and after bariatric surgery, and after a low-calorie diet intervention (data from https://adiposetissue.org/ ). Statistical analysis was performed using a two-tailed unpaired Student's t-test. Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; ns = not significant.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST and incubated overnight at 4 °C with the following primary antibodies: ANKRD53 (Proteintech, 24283-1-AP), Tubulin (Sigma–Aldrich, T6199), phospho-HSL (Ser660) (Cell Signaling Technology, 45804), HSL (Cell Signaling Technology, 4107), Phospho-PKA Substrate (Cell Signaling Technology, 9624), and Histone H3 (Cell Signaling Technology, 4499).

Techniques: Expressing, Isolation, In Vitro, Two Tailed Test

Journal: Molecular Metabolism

Article Title: ANKRD53 is downregulated in human obesity and coordinates lipolysis with mitochondrial oxidative metabolism in adipocytes

doi: 10.1016/j.molmet.2026.102330

Figure Lengend Snippet: ANKRD53 overexpression enhances lipolysis and mitochondrial oxidative capacity in human primary adipocytes (a) Schematic timeline of the experimental protocol for human primary adipocyte differentiation. (b) Representative brightfield and BODIPY-stained images of control and ANKRD53-overexpressing (OE-ANKRD53) human primary adipocytes. (c) RT-qPCR analysis of mRNA levels of adipogenic marker genes ( PPARγ , ADIPOQ , FABP4 ) in control and OE-ANKRD53 human primary adipocytes (n = 4). (d, e) Quantification of free fatty acids (FFAs) and glycerol release levels under basal and forskolin (FSK, 5 μM) stimulation in control and OE-ANKRD53 human primary adipocytes (n = 6). (f) Western blot analysis of phosphorylated hormone-sensitive lipase (p-HSL660), total HSL and phosphorylated PKA in control and OE-ANKRD53 human primary adipocytes under basal and forskolin (FSK, 5 μM) stimulation (n = 3). (g, h) Oxygen consumption rate (OCR) measurements in control and OE-ANKRD53 human primary adipocytes (n = 10). Statistical analyses were performed using a two-tailed unpaired Student's t-test for comparisons between two groups, or one-way ANOVA with Tukey multiple comparisons test for multiple-group comparisons. Data are presented as mean ± SEM.∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; ns = not significant.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST and incubated overnight at 4 °C with the following primary antibodies: ANKRD53 (Proteintech, 24283-1-AP), Tubulin (Sigma–Aldrich, T6199), phospho-HSL (Ser660) (Cell Signaling Technology, 45804), HSL (Cell Signaling Technology, 4107), Phospho-PKA Substrate (Cell Signaling Technology, 9624), and Histone H3 (Cell Signaling Technology, 4499).

Techniques: Over Expression, Staining, Control, Quantitative RT-PCR, Marker, Western Blot, Two Tailed Test

Journal: Molecular Metabolism

Article Title: ANKRD53 is downregulated in human obesity and coordinates lipolysis with mitochondrial oxidative metabolism in adipocytes

doi: 10.1016/j.molmet.2026.102330

Figure Lengend Snippet: ANKRD53 knockdown suppresses lipolysis and mitochondrial oxidative function in human primary adipocytes (a) Representative brightfield and BODIPY-stained images of control and si-ANKRD53-treated human primary adipocytes. (b) RT-qPCR analysis of adipogenic marker genes ( PPARγ, ADIPOQ , and FABP4 ) in control and si-ANKRD53 human primary adipocytes (n = 4). (c, d) Quantification of FFAs and glycerol release under basal and fo FSK (5 μM) stimulation in control and si-ANKRD53 human primary adipocytes (n = 6). (e) Western blot analysis of p-HSL660, total HSL, and p-PKA in control and si-ANKRD53 human primary adipocytes under basal and FSK (5 μM) stimulated conditions (n = 3). (f, g) OCR measurements in control and si-ANKRD53 human primary adipocytes (n = 10). Statistical analyses were performed using a two-tailed unpaired Student's t-test for comparisons between two groups, or one-way ANOVA with Tukey multiple comparisons test for multiple-group comparisons. Data are presented as mean ± SEM.∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; ns = not significant.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST and incubated overnight at 4 °C with the following primary antibodies: ANKRD53 (Proteintech, 24283-1-AP), Tubulin (Sigma–Aldrich, T6199), phospho-HSL (Ser660) (Cell Signaling Technology, 45804), HSL (Cell Signaling Technology, 4107), Phospho-PKA Substrate (Cell Signaling Technology, 9624), and Histone H3 (Cell Signaling Technology, 4499).

Techniques: Knockdown, Staining, Control, Quantitative RT-PCR, Marker, Western Blot, Two Tailed Test

Journal: Molecular Metabolism

Article Title: ANKRD53 is downregulated in human obesity and coordinates lipolysis with mitochondrial oxidative metabolism in adipocytes

doi: 10.1016/j.molmet.2026.102330

Figure Lengend Snippet: AAV-mediated ANKRD53 overexpression in iWAT enhances adipose lipolysis in vivo (a) Schematic workflow of AAV-mediated delivery of control or ANKRD53-overexpression (oe-ANKRD53) constructs into inguinal white adipose tissue (iWAT) of mice, followed by lipolysis analysis. (b) Western blot analysis of ANKRD53 protein in iWAT from control and oe-ANKRD53 mice (n = 6). (c) Representative hematoxylin and eosin (H&E) staining of iWAT sections from control and oe-ANKRD53 mice. (d, e) Quantification of plasma FFAs and glycerol release under fed and 12-h fasting in control and oe-ANKRD53 mice (n = 12) (f, g) Quantification of plasma FFAs and glycerol release under CL316,243 (1 mg/kg) at the indicated time points in control and oe-ANKRD53 mice (n = 12) (h) Western blot analysis of p-HSL660, total HSL in iWAT from control and oe-ANKRD53 mice under basal and CL316,243 (1 mg/kg) stimulated conditions (n = 3). Statistical analyses were performed using a two-tailed unpaired Student's t-test for comparisons between two groups, or one-way ANOVA with Tukey multiple comparisons test for multiple-group comparisons. Data are presented as mean ± SEM.∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST and incubated overnight at 4 °C with the following primary antibodies: ANKRD53 (Proteintech, 24283-1-AP), Tubulin (Sigma–Aldrich, T6199), phospho-HSL (Ser660) (Cell Signaling Technology, 45804), HSL (Cell Signaling Technology, 4107), Phospho-PKA Substrate (Cell Signaling Technology, 9624), and Histone H3 (Cell Signaling Technology, 4499).

Techniques: Over Expression, In Vivo, Control, Construct, Western Blot, Staining, Clinical Proteomics, Two Tailed Test

Journal: Molecular Metabolism

Article Title: ANKRD53 is downregulated in human obesity and coordinates lipolysis with mitochondrial oxidative metabolism in adipocytes

doi: 10.1016/j.molmet.2026.102330

Figure Lengend Snippet: ANKRD53 interacts with ACSL1 and promotes its mitochondrial localization (a) Representative images of immunofluorescence for ANKRD53 (red) and nuclei stained with DAPI (blue) in human primary adipocytes. (b) Western blot analysis of ANKRD53 protein in the cytoplasm fraction and nuclear fractions of human primary adipocytes. HSP90 and Histone H3 serve as cytosolic and nuclear markers, respectively (n = 2). (c) Schematic workflow of label-free proteomic analysis comparing control and OE-ANKRD53 human primary adipocytes (n = 3). (d) Schematic workflow of immunoprecipitation (IP) using FLAG-tagged ANKRD53 followed by LC-MS/MS in differentiated human primary adipocytes. (e) KEGG pathway enrichment analysis of 423 proteins upregulated in OE-ANKRD53 adipocytes (c), and ANKRD53-interacting proteins identified by IP-MS in (d). (f) Venn diagram of the overlap between the 423 upregulated proteins in OE-ANKRD53 adipocytes in (c) and 206 ANKRD53-interacting proteins identified by IP-MS in (d). (g) Co-immunoprecipitation (Co-IP) of endogenous ACSL1 in ANKRD53-overexpressing human primary adipocytes. (h) Co-IP of endogenous ANKRD53 using anti-ACSL1 antibody in human primary adipocytes. (i) Representative images of immunofluorescence for ANKRD53 (red) and ACSL1 (green) in human primary adipocytes, with nuclei stained by DAPI (blue). (j) Western blot analysis of ACSL1 in control and OE-ANKRD53 human primary adipocytes (n = 4). (k) Western blot analysis of ACSL1 protein in the cytoplasm fraction and mitochondrial fractions in control and OE-ANKRD53 human primary adipocytes. Tubulin and VDAC serve as cytosolic and mitochondrial markers, respectively (n = 3).

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST and incubated overnight at 4 °C with the following primary antibodies: ANKRD53 (Proteintech, 24283-1-AP), Tubulin (Sigma–Aldrich, T6199), phospho-HSL (Ser660) (Cell Signaling Technology, 45804), HSL (Cell Signaling Technology, 4107), Phospho-PKA Substrate (Cell Signaling Technology, 9624), and Histone H3 (Cell Signaling Technology, 4499).

Techniques: Immunofluorescence, Staining, Western Blot, Control, Immunoprecipitation, Liquid Chromatography with Mass Spectroscopy, Protein-Protein interactions, Co-Immunoprecipitation Assay

Journal: Molecular Metabolism

Article Title: ANKRD53 is downregulated in human obesity and coordinates lipolysis with mitochondrial oxidative metabolism in adipocytes

doi: 10.1016/j.molmet.2026.102330

Figure Lengend Snippet: ACSL1 knockdown partially attenuates ANKRD53-induced lipolysis and mitochondrial oxidative capacity in human primary adipocytes (a, b) Quantification of FFAs and glycerol release under FSK (5 μM) stimulation in control, OE-ANKRD53, ACSL1-knockdown, and combined OE-ANKRD53 with ACSL1-knockdown human primary adipocytes (n = 6). (c, d) OCR measurements in the four experimental groups described above (n = 10). (e) A schematic model of ANKRD53 coordinates lipolysis with mitochondrial oxidative metabolism in human adipocytes. The graph model was created with biorender.com. Statistical analysis was performed using one-way ANOVA with Tukey multiple comparisons test. Data are presented as mean ± SEM.∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns = not significant.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST and incubated overnight at 4 °C with the following primary antibodies: ANKRD53 (Proteintech, 24283-1-AP), Tubulin (Sigma–Aldrich, T6199), phospho-HSL (Ser660) (Cell Signaling Technology, 45804), HSL (Cell Signaling Technology, 4107), Phospho-PKA Substrate (Cell Signaling Technology, 9624), and Histone H3 (Cell Signaling Technology, 4499).

Techniques: Knockdown, Control