TABLE 1.

Troubleshooting table.

Step	Problem	Possible reason	Solution
5	Difficulty passing the gavage needle into the esophagus	The mouse is not positioned at the right angle, or the ball tip of the gavage needle is too large	Ensure that the main axis of the mouse is straight and the neck is extended. Use the round tip of the gavage needle to move the tongue forward. Change the type of the gavage needle to a smaller ball tip
6	Gavage solution leaks back into the oral cavity	Incorrect position of the gavage needle	The injection should only be performed while the gavage needle remains in the correct position and the tip is placed in the distal esophagus
9	Mice become weak or show signs of compromised health status	The administered CCl4 dose causes unacceptable toxicity to mice	Reduce the dose of CCl4
17	Difficulty in detaching HCA-1 from the cell culture dish	Overconfluent cell culture	Avoid overconfluence of HCA-1 cells in culture dishes. Increase the amount of trypsin-EDTA after washing with PBS
28	Difficulty in forming bubbles at the injection site	Needle is inserted too deeply into the liver tissue	Insert the needle at an angle more parallel to the liver surface and/or decrease the insertion depth of the needle
34	Tumor cell/Matrigel solution leaks from the injection site	Injected volume of cell solution is too large Cell solution is not condensed enough	Reduce the injection volume to 15 µl Increase the amount of Matrigel in the cell/Matrigel mixture or use Matrigel of a higher concentration
Box 1, step 2	Difficulty injecting the adeno-Cre into the tail vein	Perforation of the tail vein The diameter of the vein is small The mice are too small	Keep the angle of the injection needle to the skin of the tail very flat and go in very slowly Apply heat (infrared lamp, heating pad) and apply 70% (vol/vol) ethanol using a soaked gauze to dilate the tail veins Wait for another week to re-evaluate mouse weight and tail-vein diameter