Journal: EMBO Reports

Article Title: Liver ChREBP deficiency inhibits fructose-induced insulin resistance in pregnant mice and female offspring

doi: 10.1038/s44319-024-00121-w

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Rabbit anti- MLXIPL/ChREBP , ABclonal Technology , Cat# A7630.

Techniques: Isolation, Recombinant, Sequencing, Software, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: CDK6 inhibits de novo lipogenesis in white adipose tissues but not in the liver

doi: 10.1038/s41467-024-45294-z

Figure Lengend Snippet: a – d IP-Western. The same amount (1 µg) of non-immune rabbit IgG and IgG of CHREBP a , CDK6 b , proteins containing CDK phosphorylated consensus motifs (p-T*PXK/R) c , and AMPKα d were used to immunoprecipitate eWAT extracts and blotted with the respective antibodies for CHREBP a , CDK6 b , T*PXK/R c , and AMPKα d . HC indicates a heavy chain of IgG. e – i IP-Westerns. AMPKα, CDK6, CHREBP, p-AMPKα, or proteins containing consensus CDK phosphorylation motif (p-T*PXK/R) was immunoprecipitated from eWAT extracts and blotted with the indicated antibodies. The Input represented 50 μg extracts from WT eWAT. Three-four independent experiments were repeated with similar results.

Article Snippet: Antibodies used in this study included CDK6 (C-21, Santa Cruz), RUNX1 (Ab23980, Rabbit polyclonal antibody, Abcam), α-Tubulin (Sigma, T6199), GLUT1 (12939 S, Cell Signaling Technology-CST), GLUT4 (2213 S, CST), ACLY (4332 S, CST), FASN (8335 T, CST), p-ACC1 (3661 S, CST, S79), ACC1 (3676 S, CST), SCD1 (2438 S, CST), p-AMPKα (2535 S, CST, T172), AMPKα (2603 S, CST), CHREBP (58069 S, CST), mTOR (2983 S, CST), p-mTOR S2448 (5536 S, CST), p-PXS*P (2325 S, CST), and p-T*PXK/R (14371 S, CST).

Techniques: Western Blot, Phospho-proteomics, Immunoprecipitation

Journal: Nature Communications

Article Title: CDK6 inhibits de novo lipogenesis in white adipose tissues but not in the liver

doi: 10.1038/s41467-024-45294-z

Figure Lengend Snippet: a Schematic depiction of phosphorylated sites on CHREBP, which contain a N-terminal domain (1-251), a proline-rich domain, and a DNA binding domain bHLH/ZIP. The N-terminal contains two nuclear export signals (NES1 and NES2) (1-95), a nuclear localization signal (NLS, 158-190), and domains named glucose-sensing module (GSM 1-251) critical for glucose regulation including low glucose inhibitory domain (LID) and a glucose-response activation conserved element (GRACE). The numbers above the schematic diagram indicated the phosphorylation occurred on serine (S)/threonine (T) residues. Non-CDK6-specific phosphorylation of S196, S602, and S618 were indicated by dotted lines. b , c Schematic depiction of phosphorylated sites on AMPKα1 b and AMPKα2 c , respectively. AMPKα contains a kinase catalytic domain (KCD, 1-314 for α1, 1-312 for α2), an auto inhibitory domain (AID) (313/315-335), and a β/γ binding domain (471-548/552). CDK6-specific phosphorylation sites were indicated by amino acid residue numbers above the schematic diagrams. Thr-172 was highlighted in red.

Article Snippet: Antibodies used in this study included CDK6 (C-21, Santa Cruz), RUNX1 (Ab23980, Rabbit polyclonal antibody, Abcam), α-Tubulin (Sigma, T6199), GLUT1 (12939 S, Cell Signaling Technology-CST), GLUT4 (2213 S, CST), ACLY (4332 S, CST), FASN (8335 T, CST), p-ACC1 (3661 S, CST, S79), ACC1 (3676 S, CST), SCD1 (2438 S, CST), p-AMPKα (2535 S, CST, T172), AMPKα (2603 S, CST), CHREBP (58069 S, CST), mTOR (2983 S, CST), p-mTOR S2448 (5536 S, CST), p-PXS*P (2325 S, CST), and p-T*PXK/R (14371 S, CST).

Techniques: Binding Assay, Activation Assay, Phospho-proteomics, Residue

Journal: Nature Communications

Article Title: CDK6 inhibits de novo lipogenesis in white adipose tissues but not in the liver

doi: 10.1038/s41467-024-45294-z

Figure Lengend Snippet: a Phosphorylation of RUNX1 by CDK6 and other kinases such as CDK1 and CDK2 promotes RUNX1 proteolytic degradation , resulting in a reduction of RUNX1 recruitment to the proximal promoter regions of its target gens such as SCD1 , subsequently leading to reduced level of DNL-specific protein expression. CDK6 phosphorylates and activates AMPKα, which in turn phosphorylates CHREBP and ACC1. CDK6 also phosphorylates CHREBPβ. Hyper-phosphorylation of CHREBP leads to accumulation of CHREBP in the cytoplasm and reduced transcription of target genes , and hyper-phosphorylation of ACC1 , an important rate-limiting enzyme for synthesis of malonyl-CoA, which results in a reduction in lipid synthesis rates . b In the absence of CDK6 protein/kinase activity, RUNX1 is stabilized, DNL-specific protein expression is increased, and hypo-phosphorylation of CHREBP leads to accumulation of CHREBP in the nucleus and increased transcription of target genes , and hypo-phosphorylation of ACC1 translates into an increase in lipid synthesis rates .

Article Snippet: Antibodies used in this study included CDK6 (C-21, Santa Cruz), RUNX1 (Ab23980, Rabbit polyclonal antibody, Abcam), α-Tubulin (Sigma, T6199), GLUT1 (12939 S, Cell Signaling Technology-CST), GLUT4 (2213 S, CST), ACLY (4332 S, CST), FASN (8335 T, CST), p-ACC1 (3661 S, CST, S79), ACC1 (3676 S, CST), SCD1 (2438 S, CST), p-AMPKα (2535 S, CST, T172), AMPKα (2603 S, CST), CHREBP (58069 S, CST), mTOR (2983 S, CST), p-mTOR S2448 (5536 S, CST), p-PXS*P (2325 S, CST), and p-T*PXK/R (14371 S, CST).

Techniques: Phospho-proteomics, Expressing, Activity Assay