Overexpression of TLK1B-KD results in reduced histone H3 phosphorylation and poor condensation of metaphase chromosomes. Metaphase spreads from each cell line were prepared, and chromosomes were decorated with anti-histone H3P and FITC-conjugated secondary antiserum. Panels A, C, E are DAPI-stained chromosomes; panels B, D, F are stained with anti-histone H3P. Panels A and B are chromosomes of MM3MG controls; panels C and D are cells overexpressing wt TLK1B; panels E and F are cells expressing the KD mutant. The signal from phosphorylated H3 was analyzed from 10 metaphases and five random fields of each. The values were as follows: MM3MG, median pixel intensity 168 (+/-10.6 SEM); TLK1B-overexpressing cells, median 183 (+/-6.9 SEM); TLK1B-KD cells, median 133 (+/- 7.4 SEM).