Journal: Communications biology

Article Title: Nesfatin-1 and nesfatin-1-like peptide attenuate hepatocyte lipid accumulation and nucleobindin-1 disruption modulates lipid metabolic pathways.

doi: 10.1038/s42003-024-06314-2

Figure Lengend Snippet: Fig. 5 | Nesfatin-1 and NLP increased AMPK phosphorylation in oleic acid-treated cells. NLP and NESF-1 (0.1 nM) and quercetin (10 µM) increased the phosphorylation of AMPK in OA- induced HepG2/C3A cells after 2 h post incubation (a). Primary antibodies: polyclonal rabbit anti- phospho (Thr172)-AMPKα antibody (1:1000, RRID: AB_330330, cat no. 2531 S, Cell Signalling, USA), polyclonal rabbit anti-AMPKα antibody (1:1000, RRID: AB_330331, cat no. 2532 S, Cell Signalling, USA). The secondary antibodies: goat anti-rabbit IgG (H + L)-HRP conjugate antibody (1:5000, RRID: AB_11125142, cat no. 170-6515, Bio-Rad, USA) and goat anti-mouse IgG (H + L)- HRP conjugate antibody (1: 5000, RRID: AB_11125547, cat no. 170-6516, Bio-Rad, USA). Studying the NLP and NESF-1 mechanism of action on HepG2 to decrease lipid accumulation in HepG2 cells (b), HepG2 cells were treated with compound c (10 µM) for 1 h and then NLP/NESF-1/Quercetin were added to media for 24 h. Results for both parts presented are pooled from three independent stu- dies with duplicate for each treatment. All groups were pretreated with OA except untreated controls (a, b). All data are presented as mean ± SEM. Asterisk shows the significant difference between each group and OA control (a, b) while two asterisks show the significant difference between each group and untreated controls (b). Significance was set at P < 0.05.

Article Snippet: Primary antibodies: polyclonal rabbit antiphospho (Thr172)-AMPKα antibody (1:1000, RRID: AB_330330, cat no. 2531 S, Cell Signalling, USA), polyclonal rabbit anti-AMPKα antibody (1:1000, RRID: AB_330331, cat no. 2532 S, Cell Signalling, USA).

Techniques: Phospho-proteomics, Incubation, Control

Journal: Communications biology

Article Title: Nesfatin-1 and nesfatin-1-like peptide attenuate hepatocyte lipid accumulation and nucleobindin-1 disruption modulates lipid metabolic pathways.

doi: 10.1038/s42003-024-06314-2

Figure Lengend Snippet: Fig. 8 | NUCB1 disruption stimulated P-AMPK α/T-AMPK α ratio. The genetic disruption of nucleobindin 1 (Nucb1) increased the ratio of P-AMPK α/T-AMPK α in male mice liver (a), but not in female mice liver (b). The results presented are pooled from five liver samples per group. Asterisks show significant difference between knockout (KO) and wild-type (WT) groups. Significance was set at P < 0.05.

Article Snippet: Primary antibodies: polyclonal rabbit antiphospho (Thr172)-AMPKα antibody (1:1000, RRID: AB_330330, cat no. 2531 S, Cell Signalling, USA), polyclonal rabbit anti-AMPKα antibody (1:1000, RRID: AB_330331, cat no. 2532 S, Cell Signalling, USA).

Techniques: Disruption, Knock-Out

Journal: Heliyon

Article Title: FTO-mediated autophagy inhibition promotes non-small cell lung cancer progression by reducing the stability of SESN2 mRNA

doi: 10.1016/j.heliyon.2024.e27571

Figure Lengend Snippet: Primary and secondary antibodies.

Article Snippet: Rabbit anti-AMPK alpha antibody , 1:3000 , Proteintech, USA, 10929-2-AP.

Techniques:

Journal: Heliyon

Article Title: FTO-mediated autophagy inhibition promotes non-small cell lung cancer progression by reducing the stability of SESN2 mRNA

doi: 10.1016/j.heliyon.2024.e27571

Figure Lengend Snippet: FTO-regulated SESN2 expression affects the AMPK-mTOR signaling pathway. (A) Representative immunofluorescence images of H460 and H1975 cells overexpressing SESN2 along with FTO overexpression. Immunofluorescence demonstrated the fluorescence intensity of p62 in these cells (n = 3, one-way ANOVA). Scale bar: 100 μm. (B) Western blotting to detect p -AMPKα, AMPKα, p -mTOR, mTOR expression in H460 and H1975 cells overexpressing SESN2 along with FTO overexpression (n = 3, one-way ANOVA). Data are presented as means ± SD of three independent experiments. * P < 0.05, ** P < 0.01. Uncropped versions of B were added to Supplemental Material.

Article Snippet: Rabbit anti-AMPK alpha antibody , 1:3000 , Proteintech, USA, 10929-2-AP.

Techniques: Expressing, Immunofluorescence, Over Expression, Fluorescence, Western Blot

Journal: Heliyon

Article Title: FTO-mediated autophagy inhibition promotes non-small cell lung cancer progression by reducing the stability of SESN2 mRNA

doi: 10.1016/j.heliyon.2024.e27571

Figure Lengend Snippet: Schematic diagram of the proposed mechanism by which FTO deletion-mediated autophagy inhibits the progression of NSCLC. FTO deficiency promoted the interaction between IGF2BP1 and SESN2 mRNA, which enhanced SESN2 mRNA stability and its protein expression, thereby promoting autophagy and mitigating the malignant progression of NSCLC. Besides, loss of FTO upregulated SESN2 expression, which in turn induced autophagy through increased phosphorylation of AMPKα and negative regulation of mTOR.

Article Snippet: Rabbit anti-AMPK alpha antibody , 1:3000 , Proteintech, USA, 10929-2-AP.

Techniques: Expressing, Phospho-proteomics