JCS03461 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Subcellular localization of endogenous BAF during the cell cycle in cells fixed using two different methods. HeLa cells were fixed with either a mixture of 0.2% glutaraldehyde and 3.7% formaldehyde (A) or trichloroacetic acid (TCA) (B), and stained with affinity-purified anti-BAF antibody as described in the Materials and methods. Images in the left and middle columns are endogenous BAF (green in the merged image) and chromosomes stained with DAPI (blue in the merged images), respectively. Color images in the right column are merged images. The cell cycle stage is indicated on the left of each image. Bar, 10 μm.

• Supplemental Figure S2 -

  Fig. S2. Aging-dependent dislocalization of nuclear BAF in normal diploid cells. Subcellular localization of endogenous BAF stained by indirect immunofluorescence staining with a specific antibody against BAF in IMR-90 cells (A) and in WI-38 cells (B). Cell ages, represented as population doublings, are shown on the top of each image. Arrows and arrowheads in A and B indicate representative images of cells with ‘nucleus-positive’ BAF and ‘cytoplasm-positive’ BAF, respectively. Bar, 40 μm. (C) The proportions cells with different BAF localization patterns: Nuc>Cyto, Nuc-Cyto, Nuc<Cyto represent cells with nucleus-positive, uniform and cytoplasm-positive BAF, respectively. The data was derived from independent experiments with IMP-90 cells of PD19 (n=7), PD44 (n=7) and PD56 (n=4), and WI-38 cells of PD44 (n=6) and PD>52 (n=4).

• Supplemental Figure S3 -

  Fig. S3. S-phase synchronization causes accumulation of BAF into the nucleus. Endogenous BAF was stained in non-synchronized (control) and S-phase synchronized cells 0, 2, 4, and 6 hours after release from the S-phase block. The percentages of the cells with nucleus-positive (black bar, Nuc>Cyto), uniform (grey bar, uniform), and cytoplasm-positive (white bar, Nuc<Cyto) localization patterns are shown. (A) HT-1080 cells. The number of cells examined: 198 for control, 250 for 0 hours, 770 for 2 hours, 485 for 4 hours and 406 for 6 hours. (B) SiHa cells. The number of cells examined: 70 for control, 406 for 0 hours, 316 for 2 hours, 300 for 4 hours and 241 for 6 hours.