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. 2021 Aug 20;42(12):2069–2081. doi: 10.1038/s41401-021-00742-0

Fig. 3. EF inhibits iNOS expression and nitrite generation in LPS-stimulated RAW 264.7 cells and BMDMs.

Fig. 3

RAW264.7 cells (a–d) and BMDMs (e–h) were treated with EF at 0, 3, 10, 30, and 100 μM for 24 h. a, e The cytotoxicity of EF in RAW264.7 cells (a) and BMDMs (e) was assayed using CCK-8. RAW264.7 cells (b–d) and BMDMs (f–h) were preincubated with EF at 0, 3, 10, 30, and 100 μM for 0.5 h before stimulation with or without LPS (100 ng/mL) for 24 h. Cell lysates were collected for Western blot analysis, and supernatants were collected to detect the level of nitrite. The protein expression of iNOS (b, f) was measured, GAPDH was used as a loading control, and iNOS/GAPDH (c, g) was analyzed by ImageJ software. d, h The amount of nitrite in the supernatant was detected by the Griess Reagent System. Data shown as the mean ± SEM, n = 3. *P < 0.05; **P < 0.01; ***P < 0.001.