CA-5f is a potent late-stage macroautophagy/autophagy inhibitor via inhibiting autophagosome-lysosome fusion.

A549 cells or HUVECs were seeded into 100-mm culture dishes. When cells reached 80% confluency, they were exposed to DMSO or CA-5f for 6 h or 24 h. Then the cells were fixed, dehydrated, embedded, sectioned, and stained as previously described. Finally, the ultrathin sections of these samples were observed under a JEM-1230 transmission electron microscopy.

Four-week-old BALB/c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Lt and kept in specific pathogen-free conditions at the Experimental Animal Center in the Institute of Medicine in Zhengzhou University. Nude mice underwent subcutaneous injection of A549 cells (1 × 10^6) at the right scapula. 8 days after A549 cell implantation, mice exhibited palpable tumors, then they were randomly divided into 2 groups (each group n = 5). CA-5f was injected via caudal vein every 2 days for up to 30 days. DMSO was used as a control. From the initial injection, the tumor size was measured every 2 days. Tumor volumes were calculated as tumor length × (square of width)/2. At the end of the experiment, all mice were sacrificed and the tumors were resected immediately and photographed. The weight of each tumor was measured.

DMSO: 55 mg/mL (141.58 mM), Sonication is recommended.

H2O: Insoluble

10% DMSO+40% PEG300+5% Tween 80+45% Saline: 2 mg/mL (5.15 mM), Sonication is recommended.