Determining the “busiest” NYC subway line depends on the metric used (ridership, trains per hour, crowding, etc.), but based on the most common measure — annual ridership — the frontrunners are:
Historically and currently, the Lexington Avenue Line (4/5/6 trains) is the busiest trunk line in the system. It serves the East Side of Manhattan, a major job center, and connects with numerous transfer points.
The Flushing Line (7 train) often claims the title for highest peak crowding and most frequent service (with up to 30 trains per hour during rush hour pre-pandemic). It serves dense residential areas in Queens and connects to Grand Central.