Skip to main content
. 2021 Aug 13;478(15):3025–3046. doi: 10.1042/BCJ20210384

Figure 6. At the AWI, glycerol promotes the onset of meshwork formation and cluster fusion, but delays fusion completion.

Figure 6.

3.6 μM IAPP–0.04 μM bIAPP–0.08 μM avidin D fluorescein in PBS was incubated with 6% glycerol in a well of a 96-well plate. The well was imaged, z stacks encompassing the AWI collected for at least 15 h, maximum intensity projections of the time course and aggregate tracking performed. From the aggregate tracking, movement vectors and connectivity between aggregates were determined. In (A) are shown maximum intensity projections of relevant time points of an 15 h time course (top panels), the connectivity between aggregates (a red line depicts aggregates moving in the same direction, i.e. connectedness; and a black line depicts aggregates moving in the opposite direction, i.e. non-connectedness)(middle panels), and the movement vectors (bottom panels). The dashed box is an area that is further examined in (B). (B) shows an example of aggregate connectivity over relevant time points (top panels), and the corresponding snapshots from a maximum intensity projection (bottom panels). The numbers within the projections indicate individual clusters of connected aggregates that evolve over time, and the white lines show the clusters of connected aggregates (the projection and connectedness were superimposed and lines were drawn around connected aggregates). (C) Quantitation of the timing taken for fusion event to occur for several fusion events of three independent replicates (rep.) in presence of glycerol (glyc.) (left panel). Comparison of the average timing of fusion onset for reactions with bIAPP alone, and bIAPP with glycerol (right panel). ** P < 0.03 when compared with bIAPP alone. (D) Quantitation of the time taken for the fusion to be complete for several fusion events of three independent replicates in presence of glycerol (right panel). P < 0.009. Comparison of the average time taken for fusion completion for reactions with bIAPP alone, and bIAPP with glycerol (right panel). ** P < 0.03 when compared with bIAPP alone.