In The Rez Sisters, seven women journey from their reservation to Toronto to participate in "THE BIGGEST BINGO IN THE WORLD." Each woman harbors personal dreams of what the bingo jackpot could bring them. Annie yearns to "buy every single one of Patsy Cline's records" and "visit all the taverns and night clubs in Toronto to enjoy live music." Philomena imagines owning a new toilet that is "big, wide, and very white." Marie-Adele dreams of "the most beautiful, incredible goddamn island in the whole goddamn world." Veronique desires "the biggest stove on the reserve." Lastly, Pelajia hopes to construct "a nice paved road" in front of her home, frustrated that the "old chief" hasn't supported this ambition. Each woman's dream of wealth is linked to their desire to improve life at Wasy—or, in Marie-Adele's case, to leave "the rez" entirely.

However, when the women reach Toronto, luck is not on their side. Despite Philomena playing with twenty-seven cards, she only wins $600, and the rest return empty-handed after angrily confronting the bingo machine. (Marie-Adele does not return at all, as she dies during the bingo game.) Instead of dwelling on their bad luck, the "rez sisters" come to understand that their dreams cannot rely on chance alone and that they need to focus on what they can achieve themselves. During Marie-Adele's funeral, Pelajia reflects:

Well, sister, I guess you finally hit the big jackpot. Best bingo game we've ever been to in our lives, huh? You know, life's like that, I figure. When all is said and done, kinda silly, this business of living? But what choice do we have? When some fool of a being goes and puts us Indians plunk down in the middle of this old earth, dishes out this lot we got right now. But, I figure we gotta make the most of it while we're here. You certainly did. And I sure as hell am giving it one good try. For you. For me. For all of us. Promise. Really.

The remaining women start working towards bettering their lives on "the rez." Veronique takes on the responsibility of caring for Marie-Adele's children, Annie commits to practicing her singing to become a star, and Pelajia embraces her role on her roof, hammering away for a brighter future. As Philomena tells Pelajia early in the play, “This place is too much in your blood. You can't get rid of it. And it can't get rid of you." The literal and metaphorical journey in The Rez Sisters symbolizes the women grasping the importance of these words. The final appearance of Nanabush as Pelajia works on her roof, "dancing to the beat of the hammer, merrily and triumphantly," suggests that the sisters are moving in the right direction.

In her essay on The Rez Sisters in Books in Canada, Carol Bolt observes that audiences feel they are "part of an extraordinary, exuberant, life-affirming family" when experiencing the play. This response comes from Highway's ability to craft characters that represent the essence of friendship and community. The women, connected as sisters, half-sisters, or sisters-in-law, have long-standing relationships. Throughout the performance, they engage in a casual and familiar manner, exchanging jokes and gossip. Even those who profess to dislike each other, like Annie and Veronique, continue to communicate, recognizing that severing ties would be more harmful than accepting each other's peculiarities.

When verbal disputes occur, the women use the harshest insults they can muster: Philomena refers to Annie as a "slime"; Emily calls Annie a “slippery little slut''; Veronique labels Annie a "sick pervert"; Pelajia accuses Marie-Adele of being...

Nanabush, the "trickster," observes the play's events and holds a crucial place in many Native cultures and mythologies. "Our mythology spans thousands and thousands of years," Highway shared with Hartmut Lutz in Contemporary Challenges: Conversations with Canadian Native Authors. He described the trickster to the Globe and Mail's Conlogue as "central to our spiritual belief system. It's a link to a vast energy, or God, which most people only experience during extreme crises or near-death moments when they glimpse the spirit world." In The Rez Sisters, only Marie-Adele and Zhaboonigan can see Nanabush in his various manifestations, suggesting that Marie-Adele is "close to death" and that Zhaboonigan, despite her mental challenges, is more in tune with the spirit world than the other women. As Marie-Adele travels to Toronto, she meets Nanabush, who cautions her about her forthcoming death; however, her passage into the spirit realm (in the embrace of the Bingo Master) is portrayed as an awe-inspiring journey. By keeping Nanabush onstage throughout most of the play, Highway indicates that while the Trickster may be slipping from modern Native memory, he continues to play an integral role in their everyday lives.