Rabbit anti cd44

Succinic anhydride, b-PEI (MW 1,800), b-PEI (MW 250,000), 2-(7-Azabenzotriazol-1-yl)-N, N, N', N'-tetramethyluronium hexafluorophosphate (HATU), N-ethyldiisopropylamine (DPIEA) et.al was purchased from Adamas-beta (Shanghai, China). All of solvent were purchased from Macklin (Shanghai, China). Chloroquine Phosphate was obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies used for Western Blotting and immunofluorescence including rabbit anti-LC3B, anti-p62, anti-Pgp, goat anti-rabbit IgG (H + L) (HRP, Cora Lite 594) were obtained from Proteintech (Wuhan, China). Alexa fluor 647-labeled goat anti-rabbit IgG (H + L), rabbit anti-CD44, anti-ki67, Ad-GFP-LC3B, Lyso-Tracker Red, TUNEL Apoptosis Assay Kit, acid phosphatase assay kit et.al were purchased from Beyotime (Shanghai, China). All other reagents for western blotting and gel electrophoresis were obtained from Solarbio (Beijing, China).
Targeting human P-gp siRNA sequences:
Sense: 5’-AAGAAGGAAAAGAAACCAACUdTdT-3’;
Anti-sense: 5’-AGUUGGUUUCUUUUCCUUCUUdTdT-3’.
All of siRNA were obtained by GenePharma Co. Ltd. (Shanghai, China).

Cell spheroids were collected and fixed with 4% paraformaldehyde for 15 min. After washing thrice with PBS, cell spheroids were incubated with 2% (wt/v) BSA at 37 °C for 1 h, and then incubated with rabbit anti-CD44 (1: 100, Beyotime, China) overnight at 4 °C. After washing thrice with PBS, samples were incubated with a 1:100 dilution of anti-rabbit IgG for 1 h at room temperature. 4,6-diamino-2-phenylindole dihydrochloride (DAPI) was used to stain the nuclear DNA. Images were viewed and analyzed with CLSM (Andor, England).