Supplementary Figure 3: Recommended wound closure tools and abdominal suture procedure.

Incision closure information. a-c, Tools recommended for wound closure. a, Stevens needle holder. b, Adson forceps. c, Coated Vicryl absorbable suture. d, Tissue adhesive. e-l, Protocol utilized by our laboratory to suture the abdominal wall. The procedure is represented in a NOD-SCID mouse. e, Align the sharp end of the needle with the cranial end of the incision. The needle should be oriented such that it is perpendicular to the incision while the needle holders remain parallel to the incision. The sutures are applied from the cranial to the caudal end of the incision. f, Start to suture by inserting the needle from the outside of the muscle layer. Then, insert the needle on the opposite side of the incision from the inside of the muscle. Pull the needle and suture through the muscle layer so that ~2 cm of suture remains on the end of the suture. This small piece will be used to tie the suture. g, Tie three knots to secure the suture in place. The first knot will be double knot. h, The second and third knots will be single knots tied opposite of each other. The suture is now secured in place. i, Pierce the needle through the muscle, Then, grab the muscle on the opposite side and pierce the needle through it. Pull the suture tightly through. Repeat this step. Evenly separate the sutures until the incision is closed. j, Tie the suture closed as in (e) but, using the final loop of the suture. k, Tie one double knot followed by two single, opposite knots to end the suture. l, Gently tighten the suture and trim any excess away. The muscle layer is now fully closed. Sealed the skin closed with tissue adhesive or wound clips (not depicted). This procedure was approved by the Institutional Animal Care and Use Committee of the University of Michigan following the animal welfare recommendations by the National Institutes of Health.