<p>A) 30 µM of MPTQ treated or untreated cells were fixed after 24 hours of treatment and immunolabelled with an antibody specific for phosphorylated ATM (ser1983). Detection was done using Alexa fluor 594 labelled secondary antibodies. Nuclei were stained by DAPI. Z-stack images were captured from multiple random fields, processed and displayed as described in methods. B) Intensity of phospho-ATM was measured in cytoplasmic and nuclear compartment. Histograms represent mean±standard deviation of three independent images from two independent experiments. C) Western blot analysis of phosphorylated ATM on cytosolic and nuclear fraction of MPTQ treated or untreated neuro 2a cells. Blots were also immunoblotted with anti-GAPDH and anti histone H3 antibodies for normalization. The results indicate increased phosphorylated ATM level in nuclear compartment of MPTQ treated cells than untreated cells. D) Densitometric analysis of phosphorylated ATM bands was performed and the values are plotted as mean±standard deviation of three independent isolates. Statistical analysis was made by Student’s t-test and p value is displayed. p value ≤0.05 is considered significant.</p
