Fig. 2.

Blocking off-target nuclear localization of ssDNA-conjugated antibodies. a Neurons were stained with native or ssDNA-conjugated anti-bassoon antibody, anti-synapsin-I antibody, and DAPI. ssDNA-conjugated anti-bassoon antibody exhibited strong off-target nuclear localization (ii, green staining inside the nuclei), compared to the native antibody (i). This nuclear localization was reduced by blocking the fixed sample with non-specific (salmon sperm) DNA prior to immunostaining (iii) or when the anti-bassoon antibody used for staining was conjugated with ssPNA instead of ssDNA (iv). Scale bars: 20 μm. b Cross-correlation analysis of the IF images in a. Pearson correlation coefficient (PCC) of the bassoon channel (green in a) with the synapsin-I channel (red in a) for each image. Differences in PCC indicate changes in antibody staining patterns. Error bars represent 95% confidence intervals