Fig. 5.

NNK forms a potential β2AR‐Akt feedback loop in pancreatic cancer cells autophagy. (A) The mRNA and protein levels of β1‐adrenergic receptor (β1AR) were decreased by siRNA. Data are presented as mean ± SD of three separate experiments. **P < 0.01 by Student's t test. (B) Western blot assay to detect indicated proteins when cells treated with siβ1AR or siβ2AR 24 h after treatment. (C) Panc‐1 cells were pretreated with siβ2AR, ICI118551 (10 μm), CQ (10 μm), then received NNK (1 μm) treatment for 24 h, and immunoblotting was performed. (D) Panc‐1 (upper) and BxPC‐3 (lower) cells were treated with NNK (1 μm) for the indicated hours and cell total RNA was extracted. qRT‐PCR was performed to examine the mRNA levels of β1AR and β2AR. Data are presented as mean ± SD of three separate experiments. * p  < 0.05; **P < 0.01 compared with the control group by one‐way ANOVA test. (E) Relative protein levels of β2AR when cells were treated with NNK or ICI 118551. Data are presented as mean ± SD of three separate experiments. *P < 0.05 by one‐way ANOVA test. (F) Western blot to show protein expression levels when cells received NNK treatment at the indicated timepoints.