Polyglutamylase activity of tubulin tyrosine ligase-like 4 is negatively regulated by the never in mitosis gene A family kinase never in mitosis gene A -related kinase 5

Figure 2 Never in mitosis gene A-related kinase 5 changes the tubulin tyrosine ligase-like 4 and tubulin tyrosine ligase-like 5-mediated polyglutamylation. A: Analysis by Western blot of tubulin tyrosine ligase-like 4 and 5 in polyglutamylation in cells that stably express either never in mitosis gene A-related kinase 5 (NEK5) or NEK5-kinase-dead (catalytically inactive) or cells silenced for NEK5 expression. The graph next to the Western blot corresponds to the intensity of polyglutamylated α- and β-tubulins selected among the bands labeled by GT335 antibody which were measured and normalized to total α-tubulin. The arrows indicate α- and β-tubulins polyglutamylated used in this graph; and B: The intensity (area of the bands) of Western blot of the figure presented in “c” was measured using ImageJ software. NEK5: Never in mitosis gene A-related kinase 5; EYFP: Enhanced yellow fluorescent protein; TTLL: Tubulin tyrosine ligase-like.