Quantifying Podocyte Protein Localization

In paraffin-embedded mouse kidney sections, dynein subunits, phosphorylated SP1, and nephrin were coimmunofluorescent stained with podocyte markers (WT1, inverted formin 2). Using the Fiji ImageJ analysis tool, the media fluorescent intensity for podocyte-specific levels of nephrin, dynein subunit, and phosphorylated SP1 were quantified for comparison among mice with different treatments. Using the Colo2 plugin of the Fiji ImageJ software, nephrin-Dynll1 colocalization was quantified as the Manders overlap coefficient, as described in our previous work.^{10 (link)}