Journal: Endocrinology

Article Title: Neuropeptide Y Promotes mTORC1 to Regulate Chondrocyte Proliferation and Hypertrophy.

doi: 10.1210/endocr/bqac213

Figure Lengend Snippet: Figure 4. NPY activated mTORC1 activity in growth plate chondrocytes. (A, B) Chondrocytes were treated with 100 nM NPY for 48 hours in normal medium; expression of S6K1 phosphorylation (p-S6K1) and S6 phosphorylation (p-S6) were detected by Western blotting. (C, D) Chondrocytes were infected with LV3-NC or LV3-shNpy for 48 hours in normal medium. The expression of p-S6K1 and p-S6 were detected by western blotting. (E) Western blotting analysis of chondrocytes treated with or without 100 nM NPY. Cells were cultured in differentiation medium on day 0 and were harvested on day 4 and day 8, respectively. (F) Western blotting analysis of chondrocytes infected with LV3-NC or LV3-shNpy. A representative blot from 3 independent experiments was presented for each protein. (G) Immunohistochemistry assay of p-S6 expression in metatarsal growth plate. Higher magnification is shown below. Scale bars, 100 μm. The percentages of p-S6 positive cells were quantitatively analyzed (n = 6/group). *P < .05, **P < .01, ***P < .001. Data are presented as mean ± SD.

Article Snippet: Proteins were separated by 8% to 15% sodium dodecyl sulfate polyacryl amide gel electrophoresis; separated proteins were transferred onto PVDF membranes (Millipore) and were probed with the following primary antibodies: rabbit monoclonal antibodies against NPY (Cell Signaling Technology, 11976, RRID:AB_2716286), p-S6K1 (Cell Signaling Technology, 97596, RRID:AB_2800283), S6K1 (Cell Signaling Technology, 5707, RRID:AB_10 694087), pS6 (Cell Signaling Technology, 4858, RRID: AB_916156), S6 (Cell Signaling Technology, 2217, RRID: AB_331335), p-ERK (Cell Signaling Technology, 4370, RRID:AB_2315112), ERK (Cell Signaling Technology, 4695, RRID:AB_390779), TSC2 (Cell Signaling Technology, 4308, RRID:AB_10547134), p-TSC2 (Cell Signaling Technology, 40729, RRID:AB_2799183); mouse monoclonal antibodies against GAPDH (Santa Cruz Biotechnology, sc-365062, RRID:AB_10847862).

Techniques: Activity Assay, Expressing, Phospho-proteomics, Western Blot, Infection, Cell Culture, Immunohistochemistry

Journal: Endocrinology

Article Title: Neuropeptide Y Promotes mTORC1 to Regulate Chondrocyte Proliferation and Hypertrophy.

doi: 10.1210/endocr/bqac213

Figure Lengend Snippet: Figure 7. NPY/NPY2R stimulated mTORC1 activity by activation ERK1/2. (A, B) Chondrocytes were treated with or without 1 μM Y2R antagonist (Y2Rant) in the presence or absence of 100 nM NPY. The expression of S6K1 phosphorylation (p-S6K1) and S6 phosphorylation (p-S6) were detected by Western blotting. (C, D) Western blotting analysis of chondrocytes transfected with Npy2r siRNA in the presence or absence of 100 nM NPY. (E) Western blotting analysis of chondrocytes transfected with Npy2r plasmid in the presence or absence of 100 nM NPY. (F) The expression of ERK1/2 phosphorylation (p-ERK1/2) was detected by Western blotting in chondrocytes treated with or without 1 μM Y2Rant in the presence or absence of 100 nM NPY. (G) Western blotting analysis of chondrocytes transfected with Npy2r plasmid in the presence or absence of 100 nM NPY. (H) Chondrocytes were treated with or without 50 μM PD98059 (ERK1/2 inhibitor) in the presence or absence of 100 nM NPY. Levels of p-S6K1 and p-S6 were detected by Western blotting. (I) Western blotting analysis of p-ERK phosphorylated TSC2 at sites of Ser664 in chondrocytes. A representative blot from 3 independent experiments was presented for each protein. *P < .05, **P < .01, ***P < .001. Data are presented as mean ± SD.

Article Snippet: Proteins were separated by 8% to 15% sodium dodecyl sulfate polyacryl amide gel electrophoresis; separated proteins were transferred onto PVDF membranes (Millipore) and were probed with the following primary antibodies: rabbit monoclonal antibodies against NPY (Cell Signaling Technology, 11976, RRID:AB_2716286), p-S6K1 (Cell Signaling Technology, 97596, RRID:AB_2800283), S6K1 (Cell Signaling Technology, 5707, RRID:AB_10 694087), pS6 (Cell Signaling Technology, 4858, RRID: AB_916156), S6 (Cell Signaling Technology, 2217, RRID: AB_331335), p-ERK (Cell Signaling Technology, 4370, RRID:AB_2315112), ERK (Cell Signaling Technology, 4695, RRID:AB_390779), TSC2 (Cell Signaling Technology, 4308, RRID:AB_10547134), p-TSC2 (Cell Signaling Technology, 40729, RRID:AB_2799183); mouse monoclonal antibodies against GAPDH (Santa Cruz Biotechnology, sc-365062, RRID:AB_10847862).

Techniques: Activity Assay, Activation Assay, Expressing, Phospho-proteomics, Western Blot, Transfection, Plasmid Preparation

Journal: BMC Microbiology

Article Title: In vitro characterization of lactic acid bacteria and bifidobacteria from wild and domestic pigs: probiotic potential for post-weaning piglets

doi: 10.1186/s12866-024-03711-9

Figure Lengend Snippet: Origins of selected and tested strains

Article Snippet: The genome sequences of Limosilactobacillus reuteri ( L. reuteri ) DSM 20,016, Lactobacillus amylovorus ( L. amylovorus ) DSM 20,531, Lacticaseibacillus paracasei subsp. paracasei ( L. paracasei ) ATCC 25,302, Lentilactobacillus parabuchneri ( L. parabuchneri ) DSM 5707, Lactobacillus porci ( L. porci ) KCTC 21,090 and Lactobacillus delbrueckii subsp. bulgaricus ( L. delbrueckii subsp. bulgaricus ) ATCC 11,842 were included as references for ANI analysis ( https://lpsn.dsmz.de/ ).

Techniques: Sampling