TABLE 1.

List of behavioral experiments.

Experiment	Experiment description	Treatment conditions	Behavioral tests administered
1	mRNA of adenylyl cyclases in DRG and spinal cord	Systemic morphine or saline twice daily for 5 days (15 mg/kg, sc)	N/A
2	ST034307 Mechanical and thermal antinociception	Vehicle or ST034307 (2.5–10 mg/kg, ip, 100 µl) 15 min post-morphine	Von frey and thermal paw withdrawal
3	ST034307 Escalating morphine tolerance	Systemic morphine twice daily for 5 days (10 mg/kg on day 1 increasing to 40 mg/kg by day 4, sc, 100µl) + vehicle or ST034307 (2.5–10 mg/kg, ip, 100 µl) 15 min post-morphine, vehicle or ST034307 (2.5–10 mg/kg, ip, 100 µl) on Day 5	Von frey paw withdrawal
4	AC1 knockdown via intrathecal delivery: morphine antinociception	AAV9-Adcy1-shRNA or AAV9-Scrambl-shRNA delivered intrathecally, morphine doses 5–20 mg/kg, sc, measurements taken 30 min after morphine injection	Von frey paw withdrawal
5	AC1 knockdown via intrathecal delivery: escalating morphine tolerance	AAV9-Adcy1-shRNA or AAV9-Scrambl-shRNA delivered intrathecally, Twice daily escalating injections of morphine (10 mg/kg on day 1 increasing to 40 mg/kg by day 4, sc, 100 µl)	Von frey paw withdrawal
6	AC1 knockdown via intrathecal delivery: morphine tolerance	AAV9-Adcy1-shRNA or AAV9-Scrambl-shRNA delivered intrathecally, twice daily injections of morphine for 5 days (15 mg/kg, sc)	Von frey paw withdrawal
7	AC1 knockdown via intrathecal delivery: CFA model of inflammatory pain	AAV9-Adcy1-shRNA or AAV9-Scrambl-shRNA delivered intrathecally, CFA delivered intraplantar (one hindpaw)	Before CFA: Rotarod performance test, open field testing, thermal paw withdrawal testing, Von frey paw withdrawal (baseline) after CFA: Von frey paw withdrawal. burrowing testing, nesting