JCS012955 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Expression of HP1 mRNAin various human tissues. (A) Graphic representation of compiled microarray data showing the expression of the three HP1 mRNAs in 23 human tissues: (1) bladder; (2) breast; (3) whole brain; (4) pancreas; (5) bone marrow; (6) whole blood; (7) adrenal gland; (8) ovary; (9) uterus; (10) prostate; (11) lymph node; (12) lung; (13) liver; (14) tongue; (15) salivary gland; (16) pituitary gland; (17) skin; (18) thymus; (19) thyroid gland; (20) kidney; (21) testis; (22) heart and (23) stomach. The percentile values shown in the graphs are an indication of the relative expression level of the gene of interest compared to all other genes examined on the array. Arrows and asterisks indicate the highest and lowest expression levels, respectively. (B) Cumulative data (in a tabular form) presenting the relative levels of HP1α, -β, and -γ mRNAs in the different tissues. (C) Detection of HP1 proteins in human tissues. The table summarizes morphometric data obtained after immunohistochemical staining of different tissues with anti-HP1α and anti-HP1β antibodies. +, strong staining; −, undetectable; +/−, weak staining in most cells of this type; −/+, no staining in most cells of this type; ND, non-determined.

• Supplemental Figure S2 -

  Fig. S2. Immunohistochemical detection and distribution of HP1α and HP1β in human thymus. (A) Schematic representation of thymic structure and representative Hematoxylin and Eosin-stained section of human thymus (H/E; the cartoon is from Blackburn and Manley, 2004). (B) Typical profiles of the cortical and medullar regions after immunostaining with anti-HP1α and anti-HP1β antibodies. (C) Morphometric data of the staining intensity and the proportion of immunostained figures in each type of thymic cell.

• Supplemental Figure S3 -

  Fig. S3. Distribution of various chromatin markers in human immortalized T- and B-lymphocytes. HUT-78 and RPMI-8226 cells before and after co-culture or IL-2 treatment, stained with antibodies against HP1α, HP1β, HP1γ or me₃K9-H3 (green) and propidium iodide (red).

• Supplemental Figure S4 -

  Fig. S4. Distribution of various chromatin markers in human peripheral blood lymphocytes. Non-treated (NT), phytohemagglutinin-treated (PHA), lipopolysaccharide-treated (LPS) and IL-2-treated (+IL-2) human PBLs stained with antibodies against HP1α, HP1β, HP1γ or me₃K9-H3 (green) and propidium iodide (red).

• Supplemental Figure S5 -

  Fig. S5. Expression and distribution of HP1 variants in human PBLs. (A) Panoramic view of a human PBL smear stained for HP1α (green) and HP1β (red). Arrows indicate a rare sub-population of lymphocytes that have bright HP1α spots in their nuclei before induction of cell proliferation. As could be determined by staining the specimens for chromatin and nuclear envelope proteins, these cells have a normal nuclear structure (i.e. they are not apoptotic) and constitute about 4% of the total PBLs population. (B) Graphic representation of microarray data showing the relative expression of the three HP1 variants in different mononuclear cells existing in the blood stream. Notice that expression of HP1α is highest in T-lymphocytes (CD4+ or CD8+), which constitute the major fraction of PBLs.

• Supplemental Figure S6 -

  Fig. S6. Distribution of HP1 proteins at the initial stage of secondary follicle formation. The staining pattern of HP1α and HP1β during the initial steps of the GC reaction. Note that the HP1α antibodies intensely stain the GC founder cells.