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Molecular Biology of the Cell: E03-07-0487

This article contains the following additional material:

Supplemental Figures (PDF)

  • Supplemental Figure 1 Stress bodies are not major sites of accumulation of HP1alpha and gamma. HeLa cells either unstressed (37°C) or heat-shocked 1 h at 42°C followed by 1 h at 37°C (42°C) were fixed in 4% formaldehyde. Cells were co-stained with a rabbit polyclonal antibody against hnRNP HAP, to reveal stress bodies, and either with the rat mAb MCA385 specific for HP1gamma or with the mouse mAb 3446 against HP1-alpha. Primary antibodies were revealed with FITC-conjugated anti-rat or anti-mouse and rhodamine-conjugated anti-rabbit goat antibodies. Confocal laser microscopy images of the same fields are shown.

  • Supplemental Figure 2 The steady state level of HP1 proteins is not affected by heat shock. Total cell extracts were prepared from 106 HeLa cells either unstressed, heat shock 1 h at 42°C or allowed to recover at 37°C after stress. 40 mu g of proteins were loaded onto a 12% SDS-PAGE and analysed in Western blotting according to standard procedures. Primary antibodies used were: anti-HP1alpha mouse mAb 3446 (diluted 1/1000), anti-HP1beta rat mAb MAC353 (diluted 1/500), anti-HP1gamma rat mAb MAC385 (diluted 1/500) and anti-tubulin mouse mAb DM1A (diluted 1/100.000). Secondary antibodies used were: peroxidase-conjugated goat anti-rat IgG and peroxidase-conjugated goat anti-mouse IgG (Jackson Immuno Research). Lane 1: unstressed HeLa cells. Lane 2: cells heat shocked 1 h at 42°C. Lanes 3 and 4: cells allowed to recover 1 h or 3 h at 37°C after heat shock, respectively.

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