The town of Lystra is located less than 20 miles southwest of Iconium and is the most Gentile context the apostles have encountered. When Paul sees a blind man listening to him, he tells him to stand up on his feet. The man does as he is told and begins to walk. The healing performed by Barnabas and Paul leaves the Gentile crowd amazed. They mistakenly believe that Barnabas is Zeus, the king of gods, and Paul is Hermes, the messenger of gods. This is based on a local legend that says the two gods had visited Lystra in human form before. As a result, the priests of Zeus try to offer sacrifices, believing they are in divinity's presence, leading to a chaotic situation. The apostles hear the situation, and they tear their garments, usually symbolizing grief and sadness. Paul quickly responds by affirming that there is only one God and urging them to repent. He immediately challenges their belief in multiple gods and highlights that only one God is the creator of everything, which goes against their idol worship. In his message to the Gentile crowd, Paul does not directly quote from the Bible but instead relates their way of life and shared human experiences to the truth in the Scriptures. He explains theological concepts in a manner that resonates with his audience. It is unclear if people become believers during their first visit. However, on their way back through Lystra (verse 21), they encounter some believers, including Timothy (verse 16:1), who becomes a believer through Paul's ministry.

A group of Jews from Antioch and Iconium chased the apostles to Lystra, where they almost killed Paul. The city of Pisidian Antioch is approximately 100 miles (160 km) away from Lystra, and it seems that the men from there stopped in Iconium to gather more people to form their angry mob. Paul is facing a brutal ordeal at the hands of the Jews. They incited the crowd against him and attempted to stone him to death, believing him to be the main threat. Though they left him for dead outside the city, Paul miraculously survived and continued his travels the next day. This turn of events must have been shocking and horrifying for Barnabas and the other disciples.

Luke briefly describes the visit to Derbe, the farthest point reached during the first missionary journey, noting that many disciples were made due to Paul's preaching (Acts 14:21). Despite persecution, the growth pattern remained consistent. Paul and Barnabas visit Lystra, Iconium, and Antioch churches again. Paul's message "that through many tribulations we must enter the kingdom of God" (v.22) would have been particularly meaningful considering his recent experience at Lystra. Paul and Barnabas stop in Perga to preach the gospel on their way back to Antioch. Perga was a town they briefly visited earlier on the journey before Mark decided to return home. After leaving Attalia, they head back to Antioch, where they report on the journey, specifically how God had opened a door of faith to the Gentiles. Although God's work among the Gentiles is a significant step forward, not everyone is excited about including them.