Phospho-PI3 Kinase Class III (Ser244/Ser249) Antibody - #AF7421

Figure 4. MK8722 disrupts autophagic vesicle-lysosome fusion and inhibits SNARE complexes formation. (A, B) A2780 and OV90 cells were cultured in MK (20 μM, 40 μM) or an equal volume of DMEM medium for 48 h, and the expression of autophagy upstream-related proteins (PI3K, AKT, mTOR, p-PI3K, p-AKT, and p-mTOR) was analyzed by western blotting. GAPDH was used as a loading control. Quantification and analysis of western blotting results (p-mTOR, p-PI3K, and p-AKT) were performed. (C) After transient transduction of EGFP-LC3, A2780 and OV90 cells were treated with MK (20 μM) for 48 h. Fluorescence signals were examined under confocal microscopy after staining with LysoTracker Red. Scale bar: 10 μm. (D, F) Representative images showing STX17 (green) and VAMP8 (red) of A2780 and OV90 cells after 48 h of treatment with MK (20 μM). (E, G) Quantitative analysis of the relationship between STX17 and VAMP8 in (C) and (D). Scale bar: 10 μm. Scatter plots were generated using Image Pro Plus software. Results are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001. The results represent the mean values from three independent experiments (n = 3), and representative pictures are shown.

Figure 5 TZG significantly regulated the expression of NETs formation-related indicators, and the corresponding proteins in PI3K-RAC2 signaling (A–E). Serum levels of dsDNA, IL-8, TNF-α, CitH3, and MPO detected by ELISA analyses, respectively. Samples in (A)∼(E) were obtained from the blood supernatant of rats in different groups. (F–I) The ratios of p-PI3K/PI3K and p-P47/P47, and the expression levels of RAC2 and PAD4 proteins detected by western blot analyses, respectively. Samples in (F)∼(I) were obtained from MTrPs tissues of rats in different groups. The number of rats in each group was 8. Data are expressed as the mean ± S.D. “∗,” “∗∗,” and “∗∗∗,” p < 0.05, p < 0.01, and p < 0.001, respectively, comparison with the normal control group. “#,” “##,” and “###,” p < 0.05, p < 0.01, and p < 0.001, respectively, comparison with the MPS model group.

Fig. 3. Effect of FSCR on the NLRP3 inflammasome-related autophagy signaling pathway in uterine tissue of CPID rats. Detection of mRNA expression in NLRP3 inflammatory bodies by RT-PCR: NLRP3, CASP1, ASC, IL-1β, and IL-18 (A). Western blot detection of proteins in the NLRP3 inflammatory body: NLRP3, cleaved-CASP1, and cleaved-IL-1β (B). Western blot detection of proteins in the autophagy pathway: Beclin1, p-VPS34, and p62 (C). The expression of NLRP3 inflammatory bodies and autophagic vesicles in uterine tissues was observed by IF: NLRP3 was labeled with red fluorescence, VPS34 was labeled with green fluorescence, and nuclei were labeled with DAPI with blue fluorescence (D). Western blot detection of proteins in the lysosomal pathway: LAMP2, Ubiquitin and LC3 Ⅱ. (E). Results are presented as the mean ± S.E.M.