Phospho-ULK1 (Ser317) Antibody - #AF2301

Figure 2 P. bovis infection induced autophagy via activated AMPKα/ULK1 signaling in bMECs. (A) and (B) Western blot analyses of AMPKα, p-AMPKα, p-mTOR, ULK1, p-ULK1, SQSTM1/p62, Atg5 and LC3II/LC3I in bMECs. The right panel is protein quantification with ImageJ software. The bMECs were infected with P. bovis and treated with rapamycin (20 µM) for 6 hAdditionally, bMECs were pretreated with 3MA (5 mM) for 2 h, and then infected with P. bovis for 6 h Data represent means ± SD of 3 independent experiments, compared to the control group, *P < 0.05 and **P < 0.01; compared to the P. bovis group

Fig. 5. |Artesunate suppressed the excessive phosphorylation of AMPKα topromote ULK1 activation and reverse LPS tolerance.) (E-F) Tolerant PMs were stimulated with 100 ng/mL LPS alone or with Compound C (2–30 μM) for 2 h, bafilomycin A1 (20 nM)was used to evaluate the effect on autophagic flow. Phosphorylation of mTOR, AMPKα, LC3-II, and LC3-I (E) orphosphorylation sites of Ser317, 637, and 757 in ULK1 (F) were determined by western blot analysis. Both the representative blots and the densitometry data (n = 3) were shown.

Figure 7. XJZD inhibits the AMPK/ULK1(ser317) pathway. (A) The expression of ULK1 was detected by immunofluorescence assay, magnification ×400, scale bar: 20 µm (n = 6). (B) Using ImageJ software to analyse the fluorescence intensity of ULK1. (C) Representative Western blot bands of AMPK and p-AMPK. (D) Protein levels of p-AMPK/AMPK. (E) The mRNA levels of ULK1. (F) Representative Western blot bands of ULK1, p-ULK1(ser555) and p-ULK1(ser317). (G, H) Protein levels of p-ULK1(ser555)/ULK1 and p-ULK1(ser317)/ULK1 (n = 3). These data are means ± SD, ##p < 0.01 and ###p < 0.001 when compared with the control group. *p < 0.05, **p < 0.01 and ***p < 0.001 when compared with the model group. ns: no significant difference.

FIGURE 3 Ginsenoside Rh1 activates the AMPK/ULK1/FUNDC1 pathway to enhance mitochondrial autophagy in AR mice. (A) Venn diagram illustrating the overlap between AR and the targets of Ginsenoside Rh1. (B) PPI network highlighting interactions among key targets, including AMPK (PRKAG1), ULK1, and FUNDC1. (C) GO analysis of the three biological ontologies. (D) The chemical structure of Ginsenoside Rh1. (E) MD of Ginsenoside Rh1 and AMPK. (F) WB analysis demonstrating protein levels of AMPK, ULK1, and FUNDC1, as well as their phosphorylated forms. (G) WB results showing the protein levels of PINK1 and Parkin. (H, I) IF detection of PINK1 and Parkin in the nasal mucosa. Data are presented as mean ± SD (n = 8). *p