Journal: BMC biology

Article Title: Lipid metabolites affected by deficient autophagy antagonize the occurrence of autophagy through AMPK signaling in insects.

doi: 10.1186/s12915-025-02274-z

Figure Lengend Snippet: Fig. 7 Effects of metabolites on autophagy occurrence in Drosophila fat body and B. mori BmN cells. A and B LysoTracker Red staining (A), western blots and protein quantification of BmAtg8–PE and p-AMPKα (T172) (B) of Drosophila fat body at W stage after treatment with 40 μM LPE(19:0), LPC(0:0/17:0), LPC(0:0/19:0), or LPC(20:0/0:0) for 2 h, individually. C LysoTracker Red staining from nutritious and starved BmN cells after treatment with LPE(19:0), LPC(0:0/17:0), LPC(0:0/19:0), or LPC(20:0/0:0) (40 μM) for 2 h (N, normal nutrient; S, starvation). D-G Western blots and protein quantification of BmAtg8–PE and p-AMPKα (T172) after treatment with 40 μM LPE(19:0) (D), LPC(0:0/17:0) (E), LPC(0:0/19:0) (F), or LPC(20:0/0:0) (G) for 2 h in normal-nutrition and starved BmN cells (S, starvation). The solvent PBS was used as control. Data are presented as means ± SD; P values were determined by unpaired two-tailed Student’s t-test. Significance test was performed between control and each metabolite treatment. *P < 0.05, **P < 0.01, ***P < 0.001. ns, no significant difference

Article Snippet: The primary antibodies for V5 (Cell Signaling Technology, 13202S; 1:3000), FLAG (Cell Signaling Technology, 14793S; 1:3000), HA (Santa Cruz Biotechnology, sc-7392; 1:2000), BmAtg8–PE (Abcam, ab109364; 1:4000), Phospho-AMPKα (Thr172) (Cell Signaling Technology, 50081; 1:3000), and Tubulin (Beyotime Biotechnology, AT819; 1:5000) were used for western blotting according to the standard procedure as previously described [17, 41].

Techniques: Staining, Western Blot, Solvent, Control, Two Tailed Test

Journal: bioRxiv

Article Title: A negative feedback loop of the TOR signaling moderates growth and enables rapid sensing of stress signals in plants

doi: 10.1101/2020.09.06.284745

Figure Lengend Snippet: (A) Level of total and phosphorylated S6Ks (Thr-449 in S6K1 and Thr-455 in S6K2) in WT and flz8 mutants at 10 DAG. Relative level of S6K-P was quantified in comparison to total S6K (one-way ANOVA, *p ≤ 0.05, Bonferroni post-hoc test). (B) Phenotype and fresh weight of WT and mutant lines in different concentrations of TOR inhibitor AZD8055 (two-way ANOVA, *p ≤ 0.05, Bonferroni post-hoc test). (C) Sucrose-dependent hypocotyl elongation in WT and flz8.1 in response to dark treatment for 1, 2 and 3 days (two-way ANOVA, *p ≤ 0.05, Bonferroni post-hoc test).

Article Snippet: 10 DAG seedlings were used for western blotting to detect the endogenous level of the phosphorylated (Phospho-p70 S6 kinase pThr389 antibody; dilution: 1:1000; catalog no.: MA5-15117, Invitrogen) and total (Ribosomal S6 kinase 1/2 antibody; dilution: 1:2000; catalog no.: AS12 1855, Agrisera) level of S6Ks, phosphorylated (Phospho-AMPKα (Thr172) Antibody; dilution: 1:1000; catalog no.: 2531, Cell Signaling Technology) and total (SNF1-related protein kinase catalytic subunit alpha KIN10 antibody; dilution: 1:2000; catalog no.: AS10 919, Agrisera) level of SnRK1α1, and ATG8 (Autophagy-related protein ATG8 antibody; dilution: 1:2000; catalog no.: AS14 2811, Agrisera).

Techniques: Comparison, Mutagenesis

Journal: bioRxiv

Article Title: A negative feedback loop of the TOR signaling moderates growth and enables rapid sensing of stress signals in plants

doi: 10.1101/2020.09.06.284745

Figure Lengend Snippet: (A) Level of SnRK1α1 in WT and flz8 mutants at 10 DAG. Relative level of SnRK1α1 was quantified in comparison to HSP90-2 level (one-way ANOVA, *p ≤ 0.05, Bonferroni post-hoc test). (B) Level of phosphorylated SnRK1α1 (Thr-175) in WT and flz8 mutants at 10 DAG. Relative level of SnRK1α1-P was quantified in comparison to HSP90-2 level (one-way ANOVA, *p ≤ 0.05, Bonferroni post-hoc test). (C) SnRK1 activity in 10 DAG flz8 mutants in comparison to WT in AMARA peptide assay (one-way ANOVA, *p ≤ 0.05, Bonferroni post-hoc test). (D) Expression of SnRK1-activity reporter genes ( DIN6 and DIN10 ) in 10 DAG flz8 mutants in comparison to WT (one-way ANOVA, *p ≤ 0.05, Bonferroni post-hoc test). (E) Phenotype and primary root length of WT and flz8 mutants in response to ABA treatment for 5 days (two-way ANOVA, *p ≤ 0.05, Bonferroni post-hoc test).

Article Snippet: 10 DAG seedlings were used for western blotting to detect the endogenous level of the phosphorylated (Phospho-p70 S6 kinase pThr389 antibody; dilution: 1:1000; catalog no.: MA5-15117, Invitrogen) and total (Ribosomal S6 kinase 1/2 antibody; dilution: 1:2000; catalog no.: AS12 1855, Agrisera) level of S6Ks, phosphorylated (Phospho-AMPKα (Thr172) Antibody; dilution: 1:1000; catalog no.: 2531, Cell Signaling Technology) and total (SNF1-related protein kinase catalytic subunit alpha KIN10 antibody; dilution: 1:2000; catalog no.: AS10 919, Agrisera) level of SnRK1α1, and ATG8 (Autophagy-related protein ATG8 antibody; dilution: 1:2000; catalog no.: AS14 2811, Agrisera).

Techniques: Comparison, Activity Assay, Expressing