Figure 6.

Depletion of LAP2α/β and BAF by siRNA affects 3D genome organization

(A) Western blot of whole-cell extracts showing the decreased protein levels of LAP2α or BAF in cells treated with siRNA against LAP2 (siLAP2) or BAF (siBAF) or both (siLAP2-BAF) compared to scrambled siRNA (siCTRL) or untreated cells. TRF1 is unaffected, actin serves as loading control. Two independent experiments are shown.

(B) Changes in nuclear circularity 72 h post-treatment, with an increase in abnormal shapes (circularity between 0.2 and 0.6) in siRNA-treated cells compared to control. Number of analyzed cells is indicated below from at least 2 independent experiments.

(C) Relative telomere, LAD-CFHR3, LAD-CYP2C19, and LAD-CDH12 enrichment calculated over iLAD-SMIM2 or iLAD-GAPDH and control condition (set at 1) in BAF- and LAP2-depleted cells. n = 3. Mean ± SD is shown.

(D and E) Effect of double siBAF/siLAP2 on the total number of peripheral (D) and internal (E) telomeres in early G1 and G1/S phases, with a significant decrease of periphal telomeres in double-depleted siBAF/siLAP2 cells in G1/S phase. The p value of the Wilcoxon unpaired test is indicated.

(F) Distribution of the distance between each telomere and nuclear border in control (Top) and siBAF/siLAP2 treated (Bottom) nuclei in early G1 and G1/S phases indicating a decreased frequency of telomeres associated to the nuclear envelope (below dotted bar indicating 500 nm cutoff).