*2.6. Immunofluorescence Staining*

The fluorescence assay was performed as mentioned previously [26,27]. The slides were dried overnight at room temperature, washed with PBS (0.01 mM) for 8–10 min (two times), treated with proteinase K for 5 min, rinsed with PBS, and blocked with normal serum (2% goat/rabbit, as appropriate) in PBS, added with 0.1% Triton X-100. After that, the slides were incubated with primary antibodies overnight at 4 ◦C. The slides were then incubated with tetramethylrhodamine isothiocyanate–fluorescein isothiocyanate (FITC)-labeled secondary antibodies (antirabbit and antimouse, as appropriate), at room temperature for 95 min. The slides were covered using the fluorescent mounting medium. Images were taken using a confocal laser-microscope (FluoView FV 1000 MPE, Olympus, Tokyo, Japan). Integrated density was used for the quantification of the staining intensity and for the amount in the immunofluorescent microscopic image. ImageJ software (wsr@nih.gov., https://imagej.nih.gov/ij/) was used to quantify the integrated density, which represents the sum of the pixel values in an image.
