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. 2017 Mar 28;112(6):1063–1076. doi: 10.1016/j.bpj.2017.01.022

Figure 1.

Figure 1

Nuclear volume is sensitive to changes in pH, salt conditions, and ambient temperature. (A) Nuclear compartmentalization is conserved after isolation. Immunofluorescence images (midsection confocal slices) of isolated HL60 nuclei stained for DNA (Hoechst), nuclear membrane (DID), nuclear pore complex (anti-nucleoporin), lamin A/C (anti-lamin A/C), HP1α (anti-HP1α), nucleoli (NUCLEOLAR-ID Green Detection Reagent), Cajal bodies (anti-coilin), PML bodies (anti-PML), and RNA polymerase II (anti-pol II). (Yellow lines) Approximate nuclear outlines. (B) Volume versus pH (mean ± SE) for optically trapped nuclei (trap power = 0.2 W, negligible heating). n = 26, 52, 47, 58, and 43 for pH = 5, 6, 7, 8, and 9, respectively. (Insets) Phase contrast images of nuclei at low and high pH. (C) Nuclear volume versus ionic concentrations of various valencies (mean ± SE). n > 30 for each salt condition. (D) Nuclear volume measured at different ambient temperatures in 780 nm OS (trap power = 0.2 W, n = 95 and 126 for 23°C and 32°C, respectively) and a heating Petri dish (n = 98, 83 for 23 and 45°C, respectively). ∗p<0.05. Scale bars, 5 μm. To see this figure in color, go online.