Fig. 6
Oridonin attenuates ER stress by TP53/TCF4 axis in colorectal cancer. A Western blot assay showing the protein levels of TP53, Wnt, and β-catenin in colorectal cancer cells co-cultured with oridonin (0, 20, 25, 30 μM) for 24 h (n = 3). Normalized to β-actin. B Western blot assay showing the protein levels of TP53, Wnt, β-catenin, and TCF4 in RKO cells stably transfected with sg-Scb or sg-TP53 and treated with oridonin or LF3 (10 μM) (n = 3). C-E EdU staining, Tranwell, and Wound healing assays showing proliferation, invasion, and migration of RKO cells stably transfected with sg-Scb or sg-TP53 and treated with or without oridonin (n = 3). Scale bars of D: 50 μm. F Flowchart of xenografts by subcutaneous injection of RKO cells stably transfected with sg-Scb or sg-TP53 and treatment with or without oridonin. G Representative fluorescence and tumor images of xenografts by subcutaneous injection of RKO cells stably transfected with sg-Scb or sg-TP53 and treatment with or without oridonin (n = 5). H-I Representative fluorescence images (H) and the quantized histogram (I) of ER tracker-red in RKO cells stably transfected with sg-Scb or sg-TP53 and treated with or without oridonin (n = 3). Scale bars: 10 μm. J Flow cytometry analysis of intracellular ROS levels in RKO cells stably transfected with sg-Scb or sg-TP53 and treated with or without oridonin (n = 3). K The calcium indicator Fluo-3 AM detects intracellular Ca2+ levels in RKO cells stably transfected with sg-Scb or sg-TP53 and treated with or without oridonin (n = 6). The statistical results were presented as mean ± SD. Student’s t-test compared the difference in C-D and I-K, one-way ANOVA compared the difference in A, and two-way ANOVA compared the difference in E. * P < 0.05, ** P < 0.01 compared with ORI (0 μM) or sh-Scb + ORI, and # P < 0.05, ## P < 0.01 compared with sh-TP53 + DMSO. TP53: tumor protein p53