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phospho 284 acc  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho 284 acc
    Phospho 284 Acc, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 717 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phospho+acc+ser79/Phospho-Acetyl-CoA+Carboxylase+(Ser79)+Rabbit+mAb/pm41932650-133-4-27
    Average 96 stars, based on 717 article reviews
    phospho 284 acc - by Bioz Stars, 2026-09
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    Related Articles

    Blocking Assay:

    Article Title: d -Allulose Ameliorates Fructose-Induced Skeletal Muscle Insulin Resistance via Regulation of Ectopic Lipid Accumulation Independent of Body Weight Changes
    Article Snippet: .. Following the blocking step, the membranes were incubated for 1 h at room temperature with the following primary antibodies: Phospho-Akt (Ser473) #9271, AMP-activated protein kinase (AMPK)α #2532, Phospho-AMPKα (Thr172) #2531, acetyl-CoA carboxylase (ACC; C83B10) #3676, Phospho-ACC (Ser79) #3661 (all from Cell Signaling Technology, Inc., Danvers, MA, USA); and Akt1/2/3 (5C10); sc-81434 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA). .. After incubation with the primary antibodies, the membranes were washed five times for 5 min each with TBS-T and then incubated for 1 h at room temperature with a 1:1000 dilution of either horseradish peroxidase-conjugated goat anti-rabbit (Bio-Rad, Laboratories Inc., Hercules, CA, USA) or anti-mouse IgG antibody (KPL, Gaithersburg, MD, USA).

    Article Title: Preventing and correcting polycystic ovary syndrome by targeting anti-Müllerian hormone signaling in minipuberty and adulthood in mice.
    Article Snippet: Insulin response measurement Basal blood glucose levels were measured after a 6-h fast for the ITT before i.p. insulin administration (0.5 UI kg–1 body weight, Eli Lilly and Company, catalogue n HI0210) and 15, 30, 45, 60, 90 and 120 min after glucose administration using a Accu-Chek Guide glucometer. .. Tissues, all dissected during the diestrous stage, were homogenized using Parcellys 24 touch homogenizer (Bertin Technologies) in cold RIPA buffer (200 mM Tris/HCl pH 7.4, 130 mM NaCl, 10% (v/v) glycerol, 0.1% (v/v) SDS, 1% (v/v) Triton X-100, 10 mM MgCl2) with anti-proteases and anti-phosphatases (Sigma-Aldrich), and lysates were centrifuged for 30min at 18,000g and 4 C. Liver, white adipose tissue (intra-abdominal adipose tissue), ovaries and pancreas total protein lysates were separated on SDS–polyacrylamide gel electrophoresis (SDS–PAGE), electrotransferred to polyvinylidene difluoride membranes and probed successively with the following antibodies: hormone-sensitive lipase/HSL (1:1,000; Abcam, catalogue n ab45422); phospho-HSL (Ser660) (1:1000; Cell Signaling Technology, catalogue no. 4126); ACC (1:1000; Millipore, catalogue n 04-322); phospho-Acc (Ser79) (1:1000; Cell Signaling Technology, catalogue n 3661); lipoprotein lipase (1:1000; H-53, Santa Cruz Biotechnology, catalogue n sc-32885); FAS antibody (1:5000; Abcam, catalogue n ab128870); and b-actin (1:5000; Sigma-Aldrich, catalogue n A2228) and GAPDH (1:5000: Sigma-Aldrich, Catalogue n G9545-100UL), after incubation of membranes with 5% BSA blocking buffer. .. Proteins were detected using horseradish peroxidase–conjugated secondary antibodies (Dako, catalogue n PI-2000).

    Incubation:

    Article Title: d -Allulose Ameliorates Fructose-Induced Skeletal Muscle Insulin Resistance via Regulation of Ectopic Lipid Accumulation Independent of Body Weight Changes
    Article Snippet: .. Following the blocking step, the membranes were incubated for 1 h at room temperature with the following primary antibodies: Phospho-Akt (Ser473) #9271, AMP-activated protein kinase (AMPK)α #2532, Phospho-AMPKα (Thr172) #2531, acetyl-CoA carboxylase (ACC; C83B10) #3676, Phospho-ACC (Ser79) #3661 (all from Cell Signaling Technology, Inc., Danvers, MA, USA); and Akt1/2/3 (5C10); sc-81434 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA). .. After incubation with the primary antibodies, the membranes were washed five times for 5 min each with TBS-T and then incubated for 1 h at room temperature with a 1:1000 dilution of either horseradish peroxidase-conjugated goat anti-rabbit (Bio-Rad, Laboratories Inc., Hercules, CA, USA) or anti-mouse IgG antibody (KPL, Gaithersburg, MD, USA).

    Article Title: Preventing and correcting polycystic ovary syndrome by targeting anti-Müllerian hormone signaling in minipuberty and adulthood in mice.
    Article Snippet: Insulin response measurement Basal blood glucose levels were measured after a 6-h fast for the ITT before i.p. insulin administration (0.5 UI kg–1 body weight, Eli Lilly and Company, catalogue n HI0210) and 15, 30, 45, 60, 90 and 120 min after glucose administration using a Accu-Chek Guide glucometer. .. Tissues, all dissected during the diestrous stage, were homogenized using Parcellys 24 touch homogenizer (Bertin Technologies) in cold RIPA buffer (200 mM Tris/HCl pH 7.4, 130 mM NaCl, 10% (v/v) glycerol, 0.1% (v/v) SDS, 1% (v/v) Triton X-100, 10 mM MgCl2) with anti-proteases and anti-phosphatases (Sigma-Aldrich), and lysates were centrifuged for 30min at 18,000g and 4 C. Liver, white adipose tissue (intra-abdominal adipose tissue), ovaries and pancreas total protein lysates were separated on SDS–polyacrylamide gel electrophoresis (SDS–PAGE), electrotransferred to polyvinylidene difluoride membranes and probed successively with the following antibodies: hormone-sensitive lipase/HSL (1:1,000; Abcam, catalogue n ab45422); phospho-HSL (Ser660) (1:1000; Cell Signaling Technology, catalogue no. 4126); ACC (1:1000; Millipore, catalogue n 04-322); phospho-Acc (Ser79) (1:1000; Cell Signaling Technology, catalogue n 3661); lipoprotein lipase (1:1000; H-53, Santa Cruz Biotechnology, catalogue n sc-32885); FAS antibody (1:5000; Abcam, catalogue n ab128870); and b-actin (1:5000; Sigma-Aldrich, catalogue n A2228) and GAPDH (1:5000: Sigma-Aldrich, Catalogue n G9545-100UL), after incubation of membranes with 5% BSA blocking buffer. .. Proteins were detected using horseradish peroxidase–conjugated secondary antibodies (Dako, catalogue n PI-2000).

    Article Title: Timing of glucose intake drives distinct hepatic outcomes: Divergent glucose and lipid metabolism.
    Article Snippet: Objective: Excessive sugar intake is strongly associated with insulin resistance and type 2 diabetes (T2D), yet the metabolic consequences of nutrient timing—specifically glucose consumption during fasting versus feeding—remain poorly understood.. Methods: C57BL/6 mice were subjected to an every-other-day fasting (EODF) regimen and randomly divided into three groups: control (tap water only), food-glucose (FG; glucose water during feeding), and starvation-glucose (SG; glucose water during fasting).. After 22 weeks, metabolic phenotypes, hepatic lipid profiles, insulin signaling markers, and hepatic transcriptomes were analyzed.

    other:

    Article Title: Acetyl-CoA carboxylase obstructs CD8 + T cell lipid utilization in the tumor microenvironment
    Article Snippet: Phospho ACC-Ser79 (anti-human/mouse, rabbit mAb) , Cell Signaling Technology , Cat# 11818; RRID: AB_2687505.

    Article Title: Acetyl-CoA carboxylase obstructs CD8 + T cell lipid utilization in the tumor microenvironment
    Article Snippet: phospho-ACC (Ser79) (anti-mouse/human) , Cell Signaling Technology , Cat# 11818; RRID: AB_2687505.

    Nucleic Acid Electrophoresis:

    Article Title: Preventing and correcting polycystic ovary syndrome by targeting anti-Müllerian hormone signaling in minipuberty and adulthood in mice.
    Article Snippet: Insulin response measurement Basal blood glucose levels were measured after a 6-h fast for the ITT before i.p. insulin administration (0.5 UI kg–1 body weight, Eli Lilly and Company, catalogue n HI0210) and 15, 30, 45, 60, 90 and 120 min after glucose administration using a Accu-Chek Guide glucometer. .. Tissues, all dissected during the diestrous stage, were homogenized using Parcellys 24 touch homogenizer (Bertin Technologies) in cold RIPA buffer (200 mM Tris/HCl pH 7.4, 130 mM NaCl, 10% (v/v) glycerol, 0.1% (v/v) SDS, 1% (v/v) Triton X-100, 10 mM MgCl2) with anti-proteases and anti-phosphatases (Sigma-Aldrich), and lysates were centrifuged for 30min at 18,000g and 4 C. Liver, white adipose tissue (intra-abdominal adipose tissue), ovaries and pancreas total protein lysates were separated on SDS–polyacrylamide gel electrophoresis (SDS–PAGE), electrotransferred to polyvinylidene difluoride membranes and probed successively with the following antibodies: hormone-sensitive lipase/HSL (1:1,000; Abcam, catalogue n ab45422); phospho-HSL (Ser660) (1:1000; Cell Signaling Technology, catalogue no. 4126); ACC (1:1000; Millipore, catalogue n 04-322); phospho-Acc (Ser79) (1:1000; Cell Signaling Technology, catalogue n 3661); lipoprotein lipase (1:1000; H-53, Santa Cruz Biotechnology, catalogue n sc-32885); FAS antibody (1:5000; Abcam, catalogue n ab128870); and b-actin (1:5000; Sigma-Aldrich, catalogue n A2228) and GAPDH (1:5000: Sigma-Aldrich, Catalogue n G9545-100UL), after incubation of membranes with 5% BSA blocking buffer. .. Proteins were detected using horseradish peroxidase–conjugated secondary antibodies (Dako, catalogue n PI-2000).

    Pyrolysis Gas Chromatography:

    Article Title: 1,1-Diethoxyethane enhances aerobic respiration in human mitochondria via activation of AMP-activated protein kinase.
    Article Snippet: Mitotracker® Red CMXRos (#9082) and protease/phosphatase inhibitor cocktail (100X; #5872) were obtained from Cell Signaling Technology (Danvers, MA, USA). .. The following antibodies were used: AMPK (1:1000; #2532), phospho-AMPK (Thr172) (1:1000; #2535), ACC (1:1000; #3676), phospho-ACC (Ser79) (1:1000; #11818), PFKFB2 (1:1000; #13029), phosphoPFKFB2 (Ser483) (1:1000; #13064), PGC-1α (1:1000; #2178), NRF1 (1:1000; #46743), NRF2 (1:1000; #12721), β-actin (1:1000; #4970), anti-rabit IgG (1:5000; #7074) from Cell Signaling Technology (Danvers, MA, USA). .. Additional antibodies included PGC-1α (1:1000; #PA5-72948), phospho-PFKFB2 (Ser466) (1:1000; #PA5-64738), and TFAM (1:1000; #MA5-16148) from Invitrogen (Carlsbad, CA, USA).

    Article Title: 1,1-Diethoxyethane enhances aerobic respiration in human mitochondria via activation of AMP-activated protein kinase
    Article Snippet: Mitotracker® Red CMXRos (#9082) and protease/phosphatase inhibitor cocktail (100X; #5872) were obtained from Cell Signaling Technology (Danvers, MA, USA). .. The following antibodies were used: AMPK (1:1000; #2532), phospho-AMPK (Thr172) (1:1000; #2535), ACC (1:1000; #3676), phospho-ACC (Ser79) (1:1000; #11818), PFKFB2 (1:1000; #13029), phospho-PFKFB2 (Ser483) (1:1000; #13064), PGC-1α (1:1000; #2178), NRF1 (1:1000; #46743), NRF2 (1:1000; #12721), β-actin (1:1000; #4970), anti-rabit IgG (1:5000; #7074) from Cell Signaling Technology (Danvers, MA, USA). .. Additional antibodies included PGC-1α (1:1000; #PA5-72948), phospho-PFKFB2 (Ser466) (1:1000; #PA5-64738), and TFAM (1:1000; #MA5-16148) from Invitrogen (Carlsbad, CA, USA).



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    a Schematic overview of the phosphoproteomics experiment: (1) seven-day-old maize seedlings were infected with mock, SG200, or CR- hap1 and collected at 3 dpi. Created in BioRender. Doehlemann, G. (2026) https://BioRender.com/1zmsye4 . (2) Total maize proteins were extracted and separated into total proteome and phosphoproteome fractions. (3) Protein peptides were phosphoenriched using titanium dioxide. (4) Peptides were fractionated and analyzed by mass spectrometry. (5) Identified spectra were mapped to the Z. mays genome to identify phosphorylation sites. b Venn diagram showing uniquely increased phosphorylated proteins in SG200 and CR- hap1 compared to mock. c Venn diagram showing uniquely decreased phosphorylated proteins in SG200 and CR- hap1 compared to mock. d GO enrichment analysis of proteins with increased phosphorylation in SG200 vs. CR- hap1 . Significance is indicated by -Log10( P -value), with color shading from red (high significance) to blue (low significance). e Overview of the SnRK1-dependent consensus motif at P-5 and P + 4 relative to phosphorylated Ser/Thr residues. The color scheme follows the MEME Suite’s coding for amino acids. Amino acid matching the known human AMPK consensus phosphorylation sequence is colored in pink and blue at P-2 and P + 3 positions, respectively. Thirteen proteins that perfectly match the SnRK1 consensus motif are indicated in bold. When the motif criteria were relaxed to require only one of the hydrophobic residues at either P–5 or P + 4, a total of 25 proteins matched the SnRK1 motif. Log₂FC values are color-coded: black for log₂FC > 1 or < – 1 with p < 0.05, and gray for all others. f Hap1 inhibits SnRK1-dependent phosphorylation of rACC1. A GFP-fused 57-amino acid peptide surrounding the <t>Ser79</t> phosphorylation site of rat <t>acetyl-CoA</t> <t>carboxylase</t> 1 (rACC1) was co-expressed with the indicated proteins in N. benthamiana . SnRK1 activity was assessed by detecting rACC1 phosphorylation at Ser79 (pS79 ACC) using a phospho-specific <t>Anti-P-S79</t> ACC antibody. Total rACC1 levels were detected with an anti-GFP antibody. Lane 1: expression of rACC1 alone; Lane 2: co-expression of rACC1 and SnRK1; Lane 3: co-expression of rACC1, SnRK1 and Hap1; Lane 4: co-expression of rACC1 and Hap1; Lane 5: co-expression of Hap1 and SnRK1. The positions of the molecular weight ladder are shown on the right. The experiment was repeated three times with similar results. g Quantification of SnRK1-dependent rACC1 phosphorylation in the presence or absence of Hap1. The bar graph shows the fold change of P-S79 rACC1 normalized to total rACC1 (P-ACC/ACC-total). P-ACC/ACC-total ratios were scaled to the Lane 3 condition (set to 1) to obtain fold-change values. Data represent three biological replicates, each from an independent blot. Bars show mean ± SEM. Statistical significance was determined by a paired t-test on log₂ transformed values.
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    a Schematic overview of the phosphoproteomics experiment: (1) seven-day-old maize seedlings were infected with mock, SG200, or CR- hap1 and collected at 3 dpi. Created in BioRender. Doehlemann, G. (2026) https://BioRender.com/1zmsye4 . (2) Total maize proteins were extracted and separated into total proteome and phosphoproteome fractions. (3) Protein peptides were phosphoenriched using titanium dioxide. (4) Peptides were fractionated and analyzed by mass spectrometry. (5) Identified spectra were mapped to the Z. mays genome to identify phosphorylation sites. b Venn diagram showing uniquely increased phosphorylated proteins in SG200 and CR- hap1 compared to mock. c Venn diagram showing uniquely decreased phosphorylated proteins in SG200 and CR- hap1 compared to mock. d GO enrichment analysis of proteins with increased phosphorylation in SG200 vs. CR- hap1 . Significance is indicated by -Log10( P -value), with color shading from red (high significance) to blue (low significance). e Overview of the SnRK1-dependent consensus motif at P-5 and P + 4 relative to phosphorylated Ser/Thr residues. The color scheme follows the MEME Suite’s coding for amino acids. Amino acid matching the known human AMPK consensus phosphorylation sequence is colored in pink and blue at P-2 and P + 3 positions, respectively. Thirteen proteins that perfectly match the SnRK1 consensus motif are indicated in bold. When the motif criteria were relaxed to require only one of the hydrophobic residues at either P–5 or P + 4, a total of 25 proteins matched the SnRK1 motif. Log₂FC values are color-coded: black for log₂FC > 1 or < – 1 with p < 0.05, and gray for all others. f Hap1 inhibits SnRK1-dependent phosphorylation of rACC1. A GFP-fused 57-amino acid peptide surrounding the <t>Ser79</t> phosphorylation site of rat <t>acetyl-CoA</t> <t>carboxylase</t> 1 (rACC1) was co-expressed with the indicated proteins in N. benthamiana . SnRK1 activity was assessed by detecting rACC1 phosphorylation at Ser79 (pS79 ACC) using a phospho-specific <t>Anti-P-S79</t> ACC antibody. Total rACC1 levels were detected with an anti-GFP antibody. Lane 1: expression of rACC1 alone; Lane 2: co-expression of rACC1 and SnRK1; Lane 3: co-expression of rACC1, SnRK1 and Hap1; Lane 4: co-expression of rACC1 and Hap1; Lane 5: co-expression of Hap1 and SnRK1. The positions of the molecular weight ladder are shown on the right. The experiment was repeated three times with similar results. g Quantification of SnRK1-dependent rACC1 phosphorylation in the presence or absence of Hap1. The bar graph shows the fold change of P-S79 rACC1 normalized to total rACC1 (P-ACC/ACC-total). P-ACC/ACC-total ratios were scaled to the Lane 3 condition (set to 1) to obtain fold-change values. Data represent three biological replicates, each from an independent blot. Bars show mean ± SEM. Statistical significance was determined by a paired t-test on log₂ transformed values.
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    a Schematic overview of the phosphoproteomics experiment: (1) seven-day-old maize seedlings were infected with mock, SG200, or CR- hap1 and collected at 3 dpi. Created in BioRender. Doehlemann, G. (2026) https://BioRender.com/1zmsye4 . (2) Total maize proteins were extracted and separated into total proteome and phosphoproteome fractions. (3) Protein peptides were phosphoenriched using titanium dioxide. (4) Peptides were fractionated and analyzed by mass spectrometry. (5) Identified spectra were mapped to the Z. mays genome to identify phosphorylation sites. b Venn diagram showing uniquely increased phosphorylated proteins in SG200 and CR- hap1 compared to mock. c Venn diagram showing uniquely decreased phosphorylated proteins in SG200 and CR- hap1 compared to mock. d GO enrichment analysis of proteins with increased phosphorylation in SG200 vs. CR- hap1 . Significance is indicated by -Log10( P -value), with color shading from red (high significance) to blue (low significance). e Overview of the SnRK1-dependent consensus motif at P-5 and P + 4 relative to phosphorylated Ser/Thr residues. The color scheme follows the MEME Suite’s coding for amino acids. Amino acid matching the known human AMPK consensus phosphorylation sequence is colored in pink and blue at P-2 and P + 3 positions, respectively. Thirteen proteins that perfectly match the SnRK1 consensus motif are indicated in bold. When the motif criteria were relaxed to require only one of the hydrophobic residues at either P–5 or P + 4, a total of 25 proteins matched the SnRK1 motif. Log₂FC values are color-coded: black for log₂FC > 1 or < – 1 with p < 0.05, and gray for all others. f Hap1 inhibits SnRK1-dependent phosphorylation of rACC1. A GFP-fused 57-amino acid peptide surrounding the <t>Ser79</t> phosphorylation site of rat <t>acetyl-CoA</t> <t>carboxylase</t> 1 (rACC1) was co-expressed with the indicated proteins in N. benthamiana . SnRK1 activity was assessed by detecting rACC1 phosphorylation at Ser79 (pS79 ACC) using a phospho-specific <t>Anti-P-S79</t> ACC antibody. Total rACC1 levels were detected with an anti-GFP antibody. Lane 1: expression of rACC1 alone; Lane 2: co-expression of rACC1 and SnRK1; Lane 3: co-expression of rACC1, SnRK1 and Hap1; Lane 4: co-expression of rACC1 and Hap1; Lane 5: co-expression of Hap1 and SnRK1. The positions of the molecular weight ladder are shown on the right. The experiment was repeated three times with similar results. g Quantification of SnRK1-dependent rACC1 phosphorylation in the presence or absence of Hap1. The bar graph shows the fold change of P-S79 rACC1 normalized to total rACC1 (P-ACC/ACC-total). P-ACC/ACC-total ratios were scaled to the Lane 3 condition (set to 1) to obtain fold-change values. Data represent three biological replicates, each from an independent blot. Bars show mean ± SEM. Statistical significance was determined by a paired t-test on log₂ transformed values.
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    a Schematic overview of the phosphoproteomics experiment: (1) seven-day-old maize seedlings were infected with mock, SG200, or CR- hap1 and collected at 3 dpi. Created in BioRender. Doehlemann, G. (2026) https://BioRender.com/1zmsye4 . (2) Total maize proteins were extracted and separated into total proteome and phosphoproteome fractions. (3) Protein peptides were phosphoenriched using titanium dioxide. (4) Peptides were fractionated and analyzed by mass spectrometry. (5) Identified spectra were mapped to the Z. mays genome to identify phosphorylation sites. b Venn diagram showing uniquely increased phosphorylated proteins in SG200 and CR- hap1 compared to mock. c Venn diagram showing uniquely decreased phosphorylated proteins in SG200 and CR- hap1 compared to mock. d GO enrichment analysis of proteins with increased phosphorylation in SG200 vs. CR- hap1 . Significance is indicated by -Log10( P -value), with color shading from red (high significance) to blue (low significance). e Overview of the SnRK1-dependent consensus motif at P-5 and P + 4 relative to phosphorylated Ser/Thr residues. The color scheme follows the MEME Suite’s coding for amino acids. Amino acid matching the known human AMPK consensus phosphorylation sequence is colored in pink and blue at P-2 and P + 3 positions, respectively. Thirteen proteins that perfectly match the SnRK1 consensus motif are indicated in bold. When the motif criteria were relaxed to require only one of the hydrophobic residues at either P–5 or P + 4, a total of 25 proteins matched the SnRK1 motif. Log₂FC values are color-coded: black for log₂FC > 1 or < – 1 with p < 0.05, and gray for all others. f Hap1 inhibits SnRK1-dependent phosphorylation of rACC1. A GFP-fused 57-amino acid peptide surrounding the Ser79 phosphorylation site of rat acetyl-CoA carboxylase 1 (rACC1) was co-expressed with the indicated proteins in N. benthamiana . SnRK1 activity was assessed by detecting rACC1 phosphorylation at Ser79 (pS79 ACC) using a phospho-specific Anti-P-S79 ACC antibody. Total rACC1 levels were detected with an anti-GFP antibody. Lane 1: expression of rACC1 alone; Lane 2: co-expression of rACC1 and SnRK1; Lane 3: co-expression of rACC1, SnRK1 and Hap1; Lane 4: co-expression of rACC1 and Hap1; Lane 5: co-expression of Hap1 and SnRK1. The positions of the molecular weight ladder are shown on the right. The experiment was repeated three times with similar results. g Quantification of SnRK1-dependent rACC1 phosphorylation in the presence or absence of Hap1. The bar graph shows the fold change of P-S79 rACC1 normalized to total rACC1 (P-ACC/ACC-total). P-ACC/ACC-total ratios were scaled to the Lane 3 condition (set to 1) to obtain fold-change values. Data represent three biological replicates, each from an independent blot. Bars show mean ± SEM. Statistical significance was determined by a paired t-test on log₂ transformed values.

    Journal: Nature Communications

    Article Title: Ustilago maydis disrupts carbohydrate signaling networks to induce hypertrophy in host cells

    doi: 10.1038/s41467-026-69532-8

    Figure Lengend Snippet: a Schematic overview of the phosphoproteomics experiment: (1) seven-day-old maize seedlings were infected with mock, SG200, or CR- hap1 and collected at 3 dpi. Created in BioRender. Doehlemann, G. (2026) https://BioRender.com/1zmsye4 . (2) Total maize proteins were extracted and separated into total proteome and phosphoproteome fractions. (3) Protein peptides were phosphoenriched using titanium dioxide. (4) Peptides were fractionated and analyzed by mass spectrometry. (5) Identified spectra were mapped to the Z. mays genome to identify phosphorylation sites. b Venn diagram showing uniquely increased phosphorylated proteins in SG200 and CR- hap1 compared to mock. c Venn diagram showing uniquely decreased phosphorylated proteins in SG200 and CR- hap1 compared to mock. d GO enrichment analysis of proteins with increased phosphorylation in SG200 vs. CR- hap1 . Significance is indicated by -Log10( P -value), with color shading from red (high significance) to blue (low significance). e Overview of the SnRK1-dependent consensus motif at P-5 and P + 4 relative to phosphorylated Ser/Thr residues. The color scheme follows the MEME Suite’s coding for amino acids. Amino acid matching the known human AMPK consensus phosphorylation sequence is colored in pink and blue at P-2 and P + 3 positions, respectively. Thirteen proteins that perfectly match the SnRK1 consensus motif are indicated in bold. When the motif criteria were relaxed to require only one of the hydrophobic residues at either P–5 or P + 4, a total of 25 proteins matched the SnRK1 motif. Log₂FC values are color-coded: black for log₂FC > 1 or < – 1 with p < 0.05, and gray for all others. f Hap1 inhibits SnRK1-dependent phosphorylation of rACC1. A GFP-fused 57-amino acid peptide surrounding the Ser79 phosphorylation site of rat acetyl-CoA carboxylase 1 (rACC1) was co-expressed with the indicated proteins in N. benthamiana . SnRK1 activity was assessed by detecting rACC1 phosphorylation at Ser79 (pS79 ACC) using a phospho-specific Anti-P-S79 ACC antibody. Total rACC1 levels were detected with an anti-GFP antibody. Lane 1: expression of rACC1 alone; Lane 2: co-expression of rACC1 and SnRK1; Lane 3: co-expression of rACC1, SnRK1 and Hap1; Lane 4: co-expression of rACC1 and Hap1; Lane 5: co-expression of Hap1 and SnRK1. The positions of the molecular weight ladder are shown on the right. The experiment was repeated three times with similar results. g Quantification of SnRK1-dependent rACC1 phosphorylation in the presence or absence of Hap1. The bar graph shows the fold change of P-S79 rACC1 normalized to total rACC1 (P-ACC/ACC-total). P-ACC/ACC-total ratios were scaled to the Lane 3 condition (set to 1) to obtain fold-change values. Data represent three biological replicates, each from an independent blot. Bars show mean ± SEM. Statistical significance was determined by a paired t-test on log₂ transformed values.

    Article Snippet: Phospho-Acetyl-CoA Carboxylase (Ser79) (anti-P-S79 ACC) antibody (CST, #3661, Dilution: 1:1000) was used to detect phosphorylation of ACC at serine 79.

    Techniques: Phospho-proteomics, Infection, Titanium Dioxide, Mass Spectrometry, Sequencing, Activity Assay, Expressing, Molecular Weight, Transformation Assay