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Abstract The Gulf Stream affects global climate by transporting water and heat poleward. The current’s volume transport increases markedly along the U.S. East Coast. An extensive observing program using autonomous underwater gliders provides finescale, subsurface observations of hydrography and velocity spanning more than 15° of latitude along the path of the Gulf Stream, thereby filling a 1500-km-long gap between long-term transport measurements in the Florida Strait and downstream of Cape Hatteras. Here, the glider-based observations are combined with shipboard measurements along Line W near 68°W to provide a detailed picture of the along-stream transport increase. To account for the influences of Gulf Stream curvature and adjacent circulation (e.g., corotating eddies) on transport estimates, upper- and lower-bound transports are constructed for each cross–Gulf Stream transect. The upper-bound estimate for time-averaged volume transport above 1000 m is 32.9 ± 1.2 Sv (1 Sv ≡ 106 m3 s−1) in the Florida Strait, 57.3 ± 1.9 Sv at Cape Hatteras, and 75.6 ± 4.7 Sv at Line W. Corresponding lower-bound estimates are 32.3 ± 1.1 Sv in the Florida Strait, 54.5 ± 1.7 Sv at Cape Hatteras, and 69.9 ± 4.2 Sv at Line W. Using the temperature and salinity observations from gliders and Line W, waters are divided into seven classes to investigate the properties of waters that are transported by and entrained into the Gulf Stream. Most of the increase in overall Gulf Stream volume transport above 1000 m stems from the entrainment of subthermocline waters, including upper Labrador Sea Water and Eighteen Degree Water.