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. 2019 Dec 17;41(5):650–660. doi: 10.1038/s41401-019-0328-3

Fig. 4. ASM downregulation increased eNOS/NO signaling in RAECs.

Fig. 4

RAECs were incubated with 0.3 mM PA for 24 h and then transfected with ASM siRNA, or RAECs were transfected with Ob-Rb siRNA for 48 h and then treated with 10 μM AMI and 10 μM IMI for 24 h. Then, 100 nM leptin was added for 15 min prior to the collection of the cells. Summarized data showing the effects of ASM downregulation on the ratio of p-eNOS (Ser1177)/eNOS (a), eNOS activity (b), NO release (c), Ob-Rb expression (d), and the ratios of p-STAT3/STAT3 (e) and p-Akt/Akt (f) in RAECs. *P < 0.05 vs. vehicle control (Vehl Scr); #P < 0.05 vs. the PA alone-treated group; ΔP < 0.05 vs. the Ob-Rb siRNA alone-transfected group. Representative Western blot gels and summarized data showing the ratios of p-STAT3/STAT3 and p-Akt/Akt in RAECs that were pretreated with 10 μM cryptotanshinone for 24 h and then incubated with 0.3 mM PA for 24 h (g, h) or transfected with Ob-Rb siRNA for 48 h (i, j). *P < 0.05 vs. vehicle control (Vehl Scr); #P < 0.05 vs. the SCH772984 alone-treated group. The co-immunoprecipitation assay was used to test the interaction between p-STAT3 and p-Akt in RAECs treated with PA (k) or transfected with Ob-Rb siRNA (l). The data are the means ± SEMs from three experiments.