The Tonight Show with Jay Leno featured LGBTQ+ individuals and themes incidentally, primarily through guest appearances and comedic segments. The portrayal was largely neutral, avoiding strong positive or negative arcs, and neither deeply affirming nor overtly denigrating queer identity.

The Tonight Show with Jay Leno frequently incorporated transsexual themes into its comedy, often through jokes that relied on stereotypes and misgendering. These portrayals generally presented trans identity as a source of ridicule or shock, contributing to a net negative impact by lacking affirming counter-narratives or respectful depictions.

The show frequently employed satire and comedic segments to highlight perceived hypocrisy, scandals, or absurdities within Christian institutions or among prominent Christian figures, particularly televangelists. This comedic framing, while often targeting individuals or specific behaviors rather than the faith's core tenets, contributed to an overall narrative that often presented aspects of Christian adherence or leadership as foolish or hypocritical, without significant counterbalancing positive portrayals.