acsl1  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Cardiac REDD1 alters glucose and fatty acid metabolic gene expression via an mTORC1-independent, PPARα-dependent mechanism and drives hypertrophic growth

doi: 10.64898/2026.03.16.710895

Figure Lengend Snippet: A-L. Hearts of adult (12-week-old) male Redd1 Fl/Fl and Redd1 Fl/Fl, ⍺MHC-Cre mice were excised and subjected to RNA-sequencing. Genes involved in fatty acid catabolism are graphed. n=3,3 ( Cd36 , Fabp4 , Acsl1 , Acsl5 , Acss2 , Cpt2 , Acadm , Hadh , Hadhb , Decr1 , Eci1 , Acat1 ), unpaired t test. Error bars represent SEM. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Following transfer to nitrocellulose membranes, anti-REDD1 (ProteinTech-10638-1-AP), - pP70S6K (T389) (Cell Signaling-9205), -P70S6K (Cell Signaling-9202), -phospho-PDHA1 (S293) (Abcam-ab92696), -phospho-PDHA1 (S300) (Sigma-AP1064), -PDHA1 (Abcam-ab110330), - PPAR⍺ (Cayman Chemical-101710), -RXR⍺ (Cell Signaling-5388), -ACSL1 (Cell Signaling-4047), or -CARP (Santa Cruz-365056) antibodies were used at a dilution of 1:1000 in 3% bovine serum albumin (BSA; Fisher Scientific) in TBS-T.

Techniques: RNA Sequencing

Journal: bioRxiv

Article Title: Cardiac REDD1 alters glucose and fatty acid metabolic gene expression via an mTORC1-independent, PPARα-dependent mechanism and drives hypertrophic growth

doi: 10.64898/2026.03.16.710895

Figure Lengend Snippet: AC16 and AC16Δ REDD1 cardiomyocytes were cultured in DMEM, no glucose supplemented with 5.5 mM glucose and vehicle or 0.1 µM GW6471 treatment for 24 hours. A-B. Total RNA was extracted and qPCR was performed for the indicated genes. n=9,9,9 ( PDK4 ), n=9,9,6 ( ACSL1 ), 2-way ANOVA. C-E. Cardiomyocytes were harvested and subjected to western blotting with the indicated antibodies. Signals were quantified with densitometry, normalized to total protein or PDH as indicated, and plotted. n=9,9,6 (pPDH (S300)), n=7,9,8 (ACSL1), 2-way ANOVA. Error bars represent SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. M = marker.

Article Snippet: Following transfer to nitrocellulose membranes, anti-REDD1 (ProteinTech-10638-1-AP), - pP70S6K (T389) (Cell Signaling-9205), -P70S6K (Cell Signaling-9202), -phospho-PDHA1 (S293) (Abcam-ab92696), -phospho-PDHA1 (S300) (Sigma-AP1064), -PDHA1 (Abcam-ab110330), - PPAR⍺ (Cayman Chemical-101710), -RXR⍺ (Cell Signaling-5388), -ACSL1 (Cell Signaling-4047), or -CARP (Santa Cruz-365056) antibodies were used at a dilution of 1:1000 in 3% bovine serum albumin (BSA; Fisher Scientific) in TBS-T.

Techniques: Cell Culture, Western Blot, Marker

Journal: bioRxiv

Article Title: Cardiac REDD1 alters glucose and fatty acid metabolic gene expression via an mTORC1-independent, PPARα-dependent mechanism and drives hypertrophic growth

doi: 10.64898/2026.03.16.710895

Figure Lengend Snippet: A-L. Hearts of adult (12-week-old) male Redd1 Fl/Fl and Redd1 Fl/Fl, ⍺MHC-Cre mice were excised and subjected to RNA-sequencing. Genes involved in fatty acid catabolism are graphed. n=3,3 ( Cd36 , Fabp4 , Acsl1 , Acsl5 , Acss2 , Cpt2 , Acadm , Hadh , Hadhb , Decr1 , Eci1 , Acat1 ), unpaired t test. Error bars represent SEM. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: 2 μg RNA was reverse transcribed to cDNA using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). qPCR was performed using TaqMan Gene Expression Assays (ThermoFisher Scientific) and the QuantStudio 3 Real-Time PCR System (ThermoFisher Scientific) for the following genes: 18S (Mm03928990_g1), Redd1 (Mm00512504_g1), PDK1 (Hs05380290_s1), Pdk1 (Rn00587598_m1), PDK2 (Hs04965351_m1), Pdk2 (Rn00446679_m1), PDK3 (Hs03878443_s1), Pdk3 (Rn01424337_m1), PDK4 (Hs01037712_m1), Pdk4 (Rn00585577_m1), PDP1 (Hs01081518_s1), Pdp1 (Rn01437077_m1), PDP2 (Hs01934174_s1), Pdp2 (Mm02526496_s1), ACSL1 (Hs00960561_m1), or Nppb (Mm01255770_g1).

Techniques: RNA Sequencing

Journal: bioRxiv

Article Title: Cardiac REDD1 alters glucose and fatty acid metabolic gene expression via an mTORC1-independent, PPARα-dependent mechanism and drives hypertrophic growth

doi: 10.64898/2026.03.16.710895

Figure Lengend Snippet: AC16 and AC16Δ REDD1 cardiomyocytes were cultured in DMEM, no glucose supplemented with 5.5 mM glucose and vehicle or 0.1 µM GW6471 treatment for 24 hours. A-B. Total RNA was extracted and qPCR was performed for the indicated genes. n=9,9,9 ( PDK4 ), n=9,9,6 ( ACSL1 ), 2-way ANOVA. C-E. Cardiomyocytes were harvested and subjected to western blotting with the indicated antibodies. Signals were quantified with densitometry, normalized to total protein or PDH as indicated, and plotted. n=9,9,6 (pPDH (S300)), n=7,9,8 (ACSL1), 2-way ANOVA. Error bars represent SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. M = marker.

Article Snippet: 2 μg RNA was reverse transcribed to cDNA using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). qPCR was performed using TaqMan Gene Expression Assays (ThermoFisher Scientific) and the QuantStudio 3 Real-Time PCR System (ThermoFisher Scientific) for the following genes: 18S (Mm03928990_g1), Redd1 (Mm00512504_g1), PDK1 (Hs05380290_s1), Pdk1 (Rn00587598_m1), PDK2 (Hs04965351_m1), Pdk2 (Rn00446679_m1), PDK3 (Hs03878443_s1), Pdk3 (Rn01424337_m1), PDK4 (Hs01037712_m1), Pdk4 (Rn00585577_m1), PDP1 (Hs01081518_s1), Pdp1 (Rn01437077_m1), PDP2 (Hs01934174_s1), Pdp2 (Mm02526496_s1), ACSL1 (Hs00960561_m1), or Nppb (Mm01255770_g1).

Techniques: Cell Culture, Western Blot, Marker

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: For endogenous Co-IP, lysates were incubated with anti-ACSL1 antibody (Proteintech, 13989-1-AP) or control IgG (Cell Signaling Technology, 2729) at 4 °C, followed by Protein A/G magnetic bead capture (MedChemExpress, HY-K0202).

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: For endogenous Co-IP, lysates were incubated with anti-ACSL1 antibody (Proteintech, 13989-1-AP) or control IgG (Cell Signaling Technology, 2729) at 4 °C, followed by Protein A/G magnetic bead capture (MedChemExpress, HY-K0202).

Techniques: Knockdown, Control

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: Cells were incubated overnight with primary antibodies against ANKRD53 (Invitrogen, MA5-25469) and ACSL1 (Proteintech, 13989-1-AP), followed by Alexa Fluor–conjugated secondary antibodies (Abcam, ab150077, goat anti-rabbit IgG H&L, Alexa Fluor 488, green; ab150116, goat anti-mouse IgG H&L, Alexa Fluor 594, red).

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: Cells were incubated overnight with primary antibodies against ANKRD53 (Invitrogen, MA5-25469) and ACSL1 (Proteintech, 13989-1-AP), followed by Alexa Fluor–conjugated secondary antibodies (Abcam, ab150077, goat anti-rabbit IgG H&L, Alexa Fluor 488, green; ab150116, goat anti-mouse IgG H&L, Alexa Fluor 594, red).

Techniques: Knockdown, Control