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. 2023 Jul 21;14(7):458. doi: 10.1038/s41419-023-05975-x

Fig. 7. Downregulation of GOLPH3 suppresses LPS-induced inflammatory response and AKT/NF-κB signaling activation in macrophages.

Fig. 7

RAW 264.7 cells were treated with LPS (1 µg/ml) for different time points as indicated. A After LPS treatment, GOLPH3 expression was examined by western blot analysis (n = 3). B The cells were transfected with Con siRNA or GOLPH3 siRNA for 24 h and the knockdown efficiency was determined by western blot analysis. C, D The cells were transfected with Con siRNA or GOLPH3 siRNA for 24 h, and subsequently treated with LPS for 8 h. The relative mRNA levels of Golph3 and pro-inflammatory mediators (Tnfα, IL-6, Mcp1, and Nos2) were determined using real-time PCR analysis. Relative mRNA expression was normalized to that of GAPDH (n = 3). E After transfection, the cells were treated with LPS for 1 h, and determined the protein expression levels of GOLPH3, p-NF-κB p65, p-IκBα, p-AKT, and β-actin (as a loading control) using western blot analysis (n = 3). F Cells were transfected with Con siRNA or GOLPH3 siRNA for 24 h and treated with LPS for 8 h, and cell lysates were analyzed by western blotting to measure GOLPH3, iNOS, and COX2 expression. The data are presented as mean ± SEM. One-way ANOVA, followed by Bonferroni’s multiple comparisons (in A, C, D, E, F) and two-tailed Student’s t-test (in B) were used, *p < 0.05 versus Con siRNA alone, and #p < 0.05 vs LPS with Con siRNA.