Figure 2

Cellular defects in patient fibroblasts and lymphoblastoid cells, reproduced in HEK cells through siRNA and expression of mutant ZBTB24. (A) Reduced cell viability (n = 8 per time period, Student’s t-test), (B) reduced proliferation (n = 8, 36 h after plating, Student’s t-test), and (C) increased apoptosis of ZBTB24 mutant fibroblasts (activated caspase 3/7 per cell viability; n = 8, Student’s t-test). (D) Abnormal spindle (α-tubulin) formation with increase of slightly broader, unfocused microtubules poles (n = 100 metaphase LCLs, Student’s t-test, view Additional file 9: Figures S3 for further images throughout the cell cycle and specifically in metaphase cells) and (E, F) strongly reduced fluorescence signals of centrosomal marker CDK5RAP2 (but not γ-tubulin) in patient lyphoblastoid cells (n = 105 metaphase cells, Student’s t-test); total γ-tubulin levels were unaltered (n = 3, Student’s t-test). Scale bar 5 μm. View Additional file 10: Figures S4 for further images throughout the cell cycle and specifically in metaphase cells. (G) Reduced cell viability (n = 8 per time period, One-way ANOVA) and (H) mitotic spindles defect in HEK cells expressing mutant (c.1222 T > G) ZBTB24. Abnormal spindle formation with slightly broader, unfocused microtubule poles in mutant cells. Scale bar 5 μm. View Additional file 11: Figure S5 for further images throughout the cell cycle, qPCR and Western blot results. (I) ZBTB24 mRNA downregulation in HEK through siRNA (qPCR, Student’s t-test) causes (J) reduced cell culture growth (n = 3 per group per time period; Student’s t-test). ns = not significant, *p < 0.05, **p < 0.01, ***p < 0.001.