Anti snail

EVs and cells were lysed with RIPA lysis buffer including protease and phosphatase inhibitors (Boyotime, Beijing, China). Protein concentrations were determined using the Pierce™ BCA Protein Assay kit (Thermo Scientific, USA). Western blots were performed as previously described [46 (link)]. The primary antibodies used in the experiments included anti-CD63 (1:500, Santa Cruz, CA, USA), anti-TSG101 (1:500, Santa Cruz, CA, USA), anti-GM-130 (1:500, Santa Cruz, CA, USA), anti-E-cadherin (1:1000, ABclonal, Wuhan, Hubei, China), anti-N-cadherin (1:1000, Affinity Biosciences, Cincinnati, OH, USA), anti-Snail (1:1000, ABclonal, Wuhan, Hubei, China), anti-α-SMA (1:1000, Cell Signaling Technology, Danvers, MA, USA), anti-PTEN (1:1000, Cell Signaling Technology, USA); anti-PI3K (1:1000, Abcam, USA), anti-Akt, anti-p-Akt (Ser473, 1:1000, Abcam, USA), anti-p-STAT6 (Y641, 1:1000, Abcam, USA), anti-p-Smad2 (1:1000, Abcam, USA), anti-GAPDH (1:1000, ABclonal, Wuhan, Hubei, China), and anti-β-actin (1:1000, Abcam, USA). ImageJ version 1.48 was used to analyze Western blot results [47 (link)].

Bufalin and 5-Fu (Fig. 1A) were purchased from Yuanye biotechnology co. LTD (Shanghai). Primary antibodies include anti-Bcl-2, anti-Mcl-1, anti-Bax, anti-P21, anti-P27, anti-MMP9 and anti-MMP2 antibodies were purchased from Abcam; anti-E-cadherin, anti-Snail, anti-CD44, anti-CD133, anti-Sox2, anti-Oct4 and anti-Nanog antibodies were purchased from ABclonal; anti-AKT, anti-p-AKT, anti-p-ERK, anti- MEK, anti-p-MEK, anti-p-P38, anti-P38, anti-p-JNK and anti-JNK antibodies were purchased from CST.