Vertical stratification is a property of forest habitats related to the differential distribution of organisms according to the variation in the conditions, from the understory to the canopy. Here, we aimed to test whether butterfly assemblages from highly disturbed forests maintain the pattern of vertical stratification. We hypothesized that degraded forests would not exhibit vertical stratification due to the low variation in the microhabitat conditions along the vertical gradient, resulting from the canopy openness. To test this, we sampled fruit-feeding butterflies with bait traps, alternately disposed between the understory and canopy of three secondary forest fragments in a very fragmented Atlantic Forest landscape, for one year. We found that the vertical strata differed in terms of species composition, with a high contribution by the nestedness component on the beta diversity spatial variation. The understory assemblages had a higher abundance and were more diverse than the upper stratum. We demonstrated that vertical stratification is maintained even in disturbed forests; however, this does not necessarily provide support for a good quality and functioning ecosystem in these habitats. The butterfly assemblages recorded here are a subset of the species pool that inhabits conserved remnants. Thus, even being represented by species commonly found in disturbed habitats, the dynamic of vertical stratification of assemblages remains.