The penultimate glaciation (marine isotope stage (MIS) 6) is considered regionally extreme compared to the last glacial maximum, in which the European ice sheets had a vast areal extent. In contrast to the last deglaciation (19–7 ka), the penultimate deglaciation (140–130 ka) hosts one of the most rapid oceanographic changes of the late Pleistocene. In this study, we reconstructed changes in the near-surface and thermocline in the central to northeast Atlantic by analyzing sediments from two Integrated Ocean Drilling Program Expedition 306 sites. Sites U1313 (41°00.6′ N, 32°57.4′ W) and U1314 (56°21.9′ N, 27°53.3′ W) were drilled on the eastern flank of the mid-Atlantic ridge and Gardar Drift of the eastern subpolar North Atlantic, respectively. We analyzed planktonic foraminiferal assemblages, ice-rafted debris (IRD), and oxygen isotopes in two planktonic foraminifers, Globigerina bulloides, and Globorotalia inflata, from MIS 6 to 5e (185–115 ka). Warmer and colder sea-surface conditions were marked by a change in the relative abundance of polar, subpolar, and transitional planktonic foraminifers. Oxygen isotopes in G. bulloides and G. inflata suggest that the thermocline deepened at the subtropical Site U1313 during MIS 6. The lack of Globorotalia inflata prevented us from profiling the mixed layer and thermocline at the subpolar Site U1314. In contrast to MIS 6, the mixed layer and thermocline were re-stratified during the last interglacial. The lack of major IRD events at both sites suggests the stability of the Laurentide ice sheet during MIS 6 compared to the subsequent glaciation. The presence of Heinrich event 11 indicates the discharge of freshwater that freshened the sea surface, resulting in mixing between the mixed layer and thermocline. Our results were placed into a broader context using published data that shed light on the sensitivity of freshwater discharge to the North Atlantic and the following changes with a transition from a penultimate glacial to an interglacial period in surface circulation.