These results were confirmed by at least two additional biological replicates

These results were confirmed by at least two additional biological replicates. the nuclear periphery and 5 is the nuclear center. Data shown represent one technical replicate (n300 cells). These data were confirmed by two additional technical replicates.(TIF) pgen.1007393.s003.tif (315K) GUID:?6A387827-345D-44C5-BF09-DCF4DD1A8232 S4 Fig: Condensin II depletion in Kc167 cells. (A) qPCR confirming efficient knockdown of Cap-H2, Rad21, and Barren in Kc167 cells. Relative mRNA levels were normalized to levels in control RNAi samples and then to Take action5c levels. Error bars were calculated across three different biological replicates. (B) IF/FISH in Kc167 cells depleted of Cap-H2, Barren, or Rad21. Heterochromatin is usually labeled with anti-H3K9me2 antibody (green), all chromosome Oligopaints are shown in magenta, and heterochromatin FISH probes (Het) labeling the AATAT, AATAG, AACAC, 359, and dodeca satellites in gray. Scale bar 3-Cyano-7-ethoxycoumarin equals 5 m. n>500 cells per condition. (C) Tukey box plot showing the mean and distribution (minus outliers) of the Rat monoclonal to CD8.The 4AM43 monoclonal reacts with the mouse CD8 molecule which expressed on most thymocytes and mature T lymphocytes Ts / c sub-group cells.CD8 is an antigen co-recepter on T cells that interacts with MHC class I on antigen-presenting cells or epithelial cells.CD8 promotes T cells activation through its association with the TRC complex and protei tyrosine kinase lck number of Het foci. Data shown are from a single biological replicate (n>500 cells each). These results were confirmed by two additional biological replicates, respectively. ***p < 0.0001; Mann-Whitney test. (D) Quantification of mitotic chromosome spreads performed after depletion of Brown (control) or Cap-H2 in Kc167 cells. 98% of control cells and 93% of Cap-H2 depleted cells showed the normal Kc167 karyotype (observe S1 Fig; p = 0.33; Fishers Exact Test). (E) Scatter plot of nuclear volume (X-axis) versus 2L CT volume or X-2L overlap volume (Y-axis) of Cap-H2 depleted cells. Chromosome 2L volume data are shown in blue, while X-2L overlap data are shown in gray. R2 values were calculated in Prism. n = 534 cells. (F) qPCR confirming efficient knockdown of Cap-H2, Cap-D3, and SMC2 in Kc167 cells. Relative mRNA levels were normalized to levels in control RNAi samples and then to Take action5c levels. (G) Oligopaints labeling chromosomes X (green), 2L (reddish), and 2R (cyan) on representative Kc167 cell nuclei depleted of Brown (control), Cap-D3, or SMC2. Dashed lines represent the nuclear edge. Scale bar equals 5 m. (H) Tukey box plot showing CT volumes after depletion of Condensin II subunits. The data shown represent one biological replicate (n400 cells per RNAi). These data were 3-Cyano-7-ethoxycoumarin confirmed by two additional biological replicates. (I) Bar graph showing common contact frequency between the X and 2L CTs after depletion of condensin II subunits. Error bars represent the standard deviation of three biological replicates (each n400 cells per RNAi). (J) Histogram showing X-2L CT overlap as a percent of X CT volume. Binned data from a single biological experiment are shown (n>400 cells per RNAi). These results were confirmed by two additional biological replicates. ***p 0.0001; Mann-Whitney test. (K) Average 2L radial position in the nucleus determined by shell analysis with five shells of equivalent volume, where shell 1 is usually closest to the nuclear periphery and 5 is the nuclear center. Averages from a single biological experiment are shown (n>400 cells per RNAi). These results were confirmed by two additional biological replicates. ***p < 0.0001; Mann-Whitney test.(TIF) pgen.1007393.s004.tif (802K) GUID:?1E6CFF3E-D019-4096-8A11-28C4842A8FE8 S5 Fig: Cap-H2 depletion in BG3 cells. (A) Oligopaints labeling chromosomes X (green), 2L (reddish), and 2R (cyan) on representative BG3 cell nuclei depleted of Brown (control), Cap-H2, or slmb. Dashed lines represent nuclear edge. Scale bar equals 5 m. n350 cells per RNAi. (B) qPCR confirming efficient knockdown 3-Cyano-7-ethoxycoumarin of Cap-H2 and SLMB in BG3 cells. Relative mRNA levels were normalized to levels in control RNAi samples and then to Take action5c levels. (C) Tukey box plot showing CT volumes after depletion of Cap-H2 and slmb in BG3 cells. The data shown represent one biological replicate (n350 cells per RNAi). These data 3-Cyano-7-ethoxycoumarin were confirmed by one additional biological replicate..