Journal: Experimental and therapeutic medicine

Article Title: Inhibition of lipid droplet accumulation by Solanum nigrum by suppressing adipogenesis and inducing lipolysis, thermogenesis and autophagy in 3T3‑L1 cells.

doi: 10.3892/etm.2023.12032

Figure Lengend Snippet: Figure 4. Effect of SNAP on thermogenesis and autophagy in 3T3‑L1 cells. (A) Experimental design. (B) Western blot analysis in 3T3‑L1 cells treated with SNAP from D0 to D8. (C) Western blot analysis of LC3‑I and ‑II in 3T3‑L1 cells treated with SNAP from D0 to D8. *P<0.05 vs. CON group. CON, control group without the sample treatment; DMI, mixture of 0.05 mM IBMX, 1 µM dexamethasone and 10 µg/ml insulin; SNAP, Solanum nigrum aerial part extracts; p, phosphorylated; AMPK, AMP‑activated protein kinase; PGC‑1α, peroxisome proliferator‑activated receptor‑γ coactivator 1α; PRDM16, PR domain‑containing 16; UCP‑1, uncoupling protein 1; LC3, microtubule‑associated protein 1A/1B‑light chain 3.

Article Snippet: The antibodies against PPARγ (#2430), CEBPα (#8178), FABP4 (#2120), adiponectin (#2789), ATGL (#2138), HSL (#4107), Perilipin‐1 (#9349), p‐AMPK (#2535), AMPK (#5831), UCP‐1 (#14670), LC3 (#2775), β‐actin (#5125), anti‐rabbit IgG, HRP‐linked antibody (#7074) and anti‐mouse IgG, HRP‐linked antibody (#7076) were purchased from Cell Signaling (Bervely, MA, USA).

Techniques: Western Blot, Control

Journal: Theriogenology

Article Title: Expression of the apelin system in the porcine pituitary during the oestrous cycle and early pregnancy and the effect of apelin on LH and FSH secretion.

doi: 10.1016/j.theriogenology.2024.09.028

Figure Lengend Snippet: Fig. 8. The effect of apelin on the Akt, MAPK/Erk1/2, and AMPK pathways in the porcine anterior pituitary (AP) cells. The expression levels of phospho-Akt (A), phospho-MAPK/Erk1/2 (B), and phospho-AMPK (C) protein determined by Western blotting analysis, in the porcine AP lobes on days 10 to 12 of the oestrous cycle. The results are expressed as the intensity signal in arbitrary density units after normalization allowed by the presence of total Akt, MAPK/Erk1/2, and AMPK for phospho-Akt, phospho-MAPK/Erk1/2, and phospho-AMPK, respectively. Left panels – densitometric analysis of Akt, MAPK/Erk1/2, and AMPK protein ex pressions. Right panels – representative immunoblots. Results are presented as means ± S.E.M. (n = 5). Bars with different superscripts are significantly different. Different capital letters indicate P < 0.05.

Article Snippet: Primary antibodies were as follows: rabbit polyclonal to Akt (diluted 1:1000; #9272, Cell Signaling Technology, USA), rabbit polyclonal to MAPK/Erk1/2 (diluted 1:1000; #9102, Cell Signaling Technology, USA), rabbit polyclonal to AMPK (diluted 1:1000; #2532, Cell Signaling Technology, USA), rabbit polyclonal to phospho-Akt (diluted 1:500; #9271, Cell Signaling Technology, USA), rabbit polyclonal to phospho-MAPK/Erk1/2 (diluted 1:1000; #9101, Cell Signaling Technology, USA), and rabbit polyclonal to phospho-AMPK (diluted 1:1000; #2531, Cell Signaling Technology, USA).

Techniques: Expressing, Western Blot

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