Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: OR2H2 Activates CAMKKβ-AMPK-Autophagy Signaling Axis and Suppresses Senescence in VK2/E6E7 Cells.

doi: 10.3390/ph16091221

Figure Lengend Snippet: Figure 4. OR2H2 activation triggers the CaMKKß–AMPK–mTORC1 signaling pathway in VK2/E6E7 cells: (A) Immunoblot results for phospho-CaMKK-ß, CaMKK-ß, and p-CaMKK-ß/CaMKK-ß ratio; (B) phospho-AMPK, AMPK, and p-AMPK/AMPK ratio; (C) phospho-mTORC1, mTORC1, and p-mTORC1/mTORC1 ratio; (D) phospho-S6K, S6K, and p-S6K/S6K ratio; (E) phospho-S6, S6, and p- S6/S6 ratio. Tests were conducted in triplicate. Ctrl denotes control (0.1% ethanol); A769662 (AMPK agonist), 100 µM; Ald 13-13 refers to aldehyde 13-13 at 500 µM; Rapa (rapamycin) is at 100 nM. Data are displayed as mean ± SD. Two-tailed unpaired t-test was used for multiple or two-group comparisons. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: Primary antibodies to β-actin (SC-47778, 1:1000), CaMKK-ß (SC-50341, 1:1000), and AMPKα1/2 (SC-25792, 1:1000) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Activation Assay, Western Blot, Control, Two Tailed Test

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: OR2H2 Activates CAMKKβ-AMPK-Autophagy Signaling Axis and Suppresses Senescence in VK2/E6E7 Cells.

doi: 10.3390/ph16091221

Figure Lengend Snippet: Figure 6. Aldehyde 13-13’s influence on the CAMKKβ–AMPK–autophagy pathway is negated following OR2H2 gene knockdown: (a) OR2H2 gene knockdown analysis in cells transfected with control (scrambled) and varying concentrations of OR2H2 siRNA; (b) Immunoblot results for phospho-CaMKK-ß, CaMKK-ß, and p-CaMKK-ß/CaMKK-ß ratio; (c) phospho-AMPK, AMPK, and p-AMPK/AMPK ratio; (d) LC3I, LC3II, and LC3II/LC3I ratio. Experiments were performed in triplicate. Ctrl, control (0.1% DMSO); Ald 13-13, aldehyde 13-13 (500 µM). Data are shown as mean ± SD. Two-tailed unpaired t-test was used for multiple or two-group comparisons. **, p < 0.01; ***, p < 0.001.

Article Snippet: Primary antibodies to β-actin (SC-47778, 1:1000), CaMKK-ß (SC-50341, 1:1000), and AMPKα1/2 (SC-25792, 1:1000) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Knockdown, Transfection, Control, Western Blot, Two Tailed Test

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Evaluation of lung protection of Sanghuangporus sanghuang through TLR4/NF-κB/MAPK, keap1/Nrf2/HO-1, CaMKK/AMPK/Sirt1, and TGF-β/SMAD3 signaling pathways mediating apoptosis and autophagy.

doi: 10.1016/j.biopha.2023.115080

Figure Lengend Snippet: Fig. 7. The protein levels of CaMKK, AMPK, and Sirt-1 in mice with bleomycin-induced pulmonary fibrosis were analyzed by Western blot after being treated with DXM and SS for 21 days. The mean ± S.E.M values (n = 3) were reported after densitometric analysis. ### p < 0.001 compared with the control group, while ***

Article Snippet: The signals were detected and analyzed using the Kodak Gel Logic 1500 Imaging Software. p-CaMKK (#12818, 1:1000), p-IκB (#2859 S, 1:1000), p-ERK (#3192, 1:1000), Beclin-1 (#3738, 1:1000), ERK (#4695, 1:1000), AMPK (#5831, 1:1000), and LC3 AB-I/II (12741 S, 1:1000) primary antibodies were purchased from Cell Signaling.

Techniques: Western Blot, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Evaluation of lung protection of Sanghuangporus sanghuang through TLR4/NF-κB/MAPK, keap1/Nrf2/HO-1, CaMKK/AMPK/Sirt1, and TGF-β/SMAD3 signaling pathways mediating apoptosis and autophagy.

doi: 10.1016/j.biopha.2023.115080

Figure Lengend Snippet: Fig. 10. A scheme displaying the protective effect of SS against bleomycin-associated lung fibrosis. An increase is denoted by the green arrows. A decrease is denoted by the red ar rows. ROS: reactive oxygen species; MAPK: mitogen-activated protein kinase; JNK: C-jun NH2-terminal kinase; ERK: extracellular-signal- regulated kinase; NF-κB: nuclear factor of κB; HO-1: heme oxygenase 1; GPx3: glutathione peroxidases 3; Nrf2: nuclear-factor-erythroid-2- related factor 2; SOD-1: superoxide dismutase 1; IκB: inhibitor of the nuclear factor kappa B; TLR-4: toll-like receptor 4; AMPK: 5′-adenosine- monophosphate-activated protein kinase; CaMKK: calcium/calmodulin-dependent pro tein kinase kinase; Sirt-1: Sirtuin-1; COX-2: cyclooxygenase-2; iNOS: inducible nitric oxide synthase; Bcl-2: B-cell lymphoma 2; p53: tumor protein p53; Bax: Bcl-2-associated X; TNF-α: tumor necrosis factor-α; IL-1ß: interleukin-1β; IL-6: interleukin-6; Smad3: mothers against decapentaplegic homolog 3; MMP: matrix met alloproteinase; α-SMA: α-smooth muscle actin.

Article Snippet: The signals were detected and analyzed using the Kodak Gel Logic 1500 Imaging Software. p-CaMKK (#12818, 1:1000), p-IκB (#2859 S, 1:1000), p-ERK (#3192, 1:1000), Beclin-1 (#3738, 1:1000), ERK (#4695, 1:1000), AMPK (#5831, 1:1000), and LC3 AB-I/II (12741 S, 1:1000) primary antibodies were purchased from Cell Signaling.

Techniques:

Journal: Cell

Article Title: Neuronal Inactivity Co-opts LTP Machinery to Drive Potassium Channel Splicing and Homeostatic Spike Widening

doi: 10.1016/j.cell.2020.05.013

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse anti-αCaMKK , Santa Cruz , Cat#sc-17827;RRID:AB_2275110.

Techniques: Recombinant, Mutagenesis, Immunoprecipitation, Isolation, Protein Extraction, Lysis, Labeling, Construct, Plasmid Preparation, shRNA, Software

Journal: Animal Nutrition

Article Title: Calcium overload and reactive oxygen species accumulation induced by selenium deficiency promote autophagy in swine small intestine

doi: 10.1016/j.aninu.2021.05.005

Figure Lengend Snippet: Gene-special primers used in the quantitative real-time PCR.

Article Snippet: The membranes were blocked with 5% skim milk for 2 h and incubated overnight with diluted primary antibodies against Beclin 1 (1:500, Wanleibio, China), LC3 (1:500, ABclonal, China), mTOR (1:500, Wanleibio, China), AMPK (1:800, the polyclonal antibody produced by our lab), CAMKK-β (1:500, Proteintech, China), SERCA (1:500, the polyclonal antibody produced by our lab) and β-actin (1:10,000, ABclonal, China) followed by goat anti-rabbit IgG (H + L) (1:10,000, Immuno Way, China).

Techniques: Sequencing

Journal: Animal Nutrition

Article Title: Calcium overload and reactive oxygen species accumulation induced by selenium deficiency promote autophagy in swine small intestine

doi: 10.1016/j.aninu.2021.05.005

Figure Lengend Snippet: Detection of autophagy-Ca 2+ homeostasis-related genes in small intestine tissue. The expression of autophagy-Ca 2+ homeostasis related genes at (A) mRNA and (B) protein levels in small intestine tissue of the control and selenium-deficient groups, respectively. The ∗ indicates significant differences (∗ P < 0.05, ∗∗ P < 0.002, ∗∗∗ P < 0.0002, ∗∗∗∗ P < 0.0001) between the 2 groups, and the data are presented as the mean ± SD ( n = 6). LC3 = microtubule-associated proteins; ATG = autophagy-related protein; mTOR = mammalian target; CAMKK-β = calmodulin-dependent protein kinase kinase β; AMPK = adenosine 5′-monophosphate-activated protein kinase; SERCA = sarco(endo)plasmic reticulum Ca 2+ -ATPase.

Article Snippet: The membranes were blocked with 5% skim milk for 2 h and incubated overnight with diluted primary antibodies against Beclin 1 (1:500, Wanleibio, China), LC3 (1:500, ABclonal, China), mTOR (1:500, Wanleibio, China), AMPK (1:800, the polyclonal antibody produced by our lab), CAMKK-β (1:500, Proteintech, China), SERCA (1:500, the polyclonal antibody produced by our lab) and β-actin (1:10,000, ABclonal, China) followed by goat anti-rabbit IgG (H + L) (1:10,000, Immuno Way, China).

Techniques: Expressing, Control

Journal: Animal Nutrition

Article Title: Calcium overload and reactive oxygen species accumulation induced by selenium deficiency promote autophagy in swine small intestine

doi: 10.1016/j.aninu.2021.05.005

Figure Lengend Snippet: Detection of autophagy-Ca 2+ homeostasis-related genes in IPEC-J2 cells. The expression of autophagy-Ca 2+ homeostasis related genes at (A) mRNA and (B) protein levels in IPEC-J2 cells of the control and selenium-deficient groups, respectively. The ∗ in indicates significant differences (∗ P < 0.05, ∗∗ P < 0.002, ∗∗∗ P < 0.0002, ∗∗∗∗ P < 0.0001) between the 2 groups, and the data are presented as the mean ± SD ( n = 6). IPEC-J2 = intestinal porcine enterocyte cell line; LC3 = microtubule-associated proteins; ATG = autophagy-related protein; mTOR = mammalian target; CAMKK = calmodulin-dependent protein kinase kinase; AMPK = adenosine 5′-monophosphate-activated protein kinase; SERCA = sarco(endo)plasmic reticulum Ca 2+ -ATPase.

Article Snippet: The membranes were blocked with 5% skim milk for 2 h and incubated overnight with diluted primary antibodies against Beclin 1 (1:500, Wanleibio, China), LC3 (1:500, ABclonal, China), mTOR (1:500, Wanleibio, China), AMPK (1:800, the polyclonal antibody produced by our lab), CAMKK-β (1:500, Proteintech, China), SERCA (1:500, the polyclonal antibody produced by our lab) and β-actin (1:10,000, ABclonal, China) followed by goat anti-rabbit IgG (H + L) (1:10,000, Immuno Way, China).

Techniques: Expressing, Control