Fig. 4

SCA14 mutations reduce PMA-induced membrane translocation of PKCγ. a Control and patient iPSCs were immunostained for PKCγ before or after treatment with PMA. The cell membrane was stained with an antibody against sodium potassium ATPase. Cell nuclei are visualized by Hoechst staining. In unstimulated control iPSCs, PKCγ was expressed as small dots in the cytoplasm (white solid arrowhead). After 5 min of PMA treatment, PKCγ was found at the plasma membrane (white hollow arrowheads), and returned to the cytoplasm after 15 min of PMA treatment (white solid arrowhead). In unstimulated SCA14 iPSCs, large aggregates (white arrowheads) of PKCγ were present in the cytoplasm. PKCγ inclusions remained in the cytoplasm (white arrowheads) throughout the treatment with PMA. Scale bar: 10 μm. b PKCγ in SCA14 iPSCs showed significantly less membrane association than in control iPSCs in response to PMA stimulation (n = 3, ****p < 0.0001, two-way ANOVA followed by Bonferroni’s post-hoc test)