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.