Then 1 µl of C/EBPβ 3'UTR RNA, or 1 µl of εV1-2 (kindly provided by Zhi-Qi Zhao), both in increasing concentrations, were added to the respective tubes.
The colloidal suspension described above was added to cell cultures of 3T3-L1 murine fibroblasts (American Type Culture Collection, Manassas, VA, USA), HSC-2 (a human oral squamous carcinoma line), and S-G (a human immortalized gingival epithelioid cell line, both kindly provided by Dr. Babich, Yeshiva University, New York).
Boolean combination gates were created and data was exported to PESTLE and SPICE for analysis (both kindly provided by Mario Roederer).
We used a clinical S. aureus strain belonging to the USA300 lineage, and its isogenic ΔlukS/F-PV derivative (LAC and LACΔpvl, respectively), both kindly provided by Frank DeLeo.
To label the visual pigments of cone and rod photoreceptors, we used antisera sc-14363 (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) and JH455 against the S opsin, JH492 against the L opsin (both kindly provided by J. Nathans), and rho4D2 against the rod opsin (kindly provided by R. S. Molday).
Polyclonal TNFα-antiserum from sheep, IFNγ antiserum from rabbit (both kindly provided by G. Tiegs, University of Erlangen), or for vehicle normal sheep or rabbit serum (Roth, Karlsruhe, Germany) were administered ip at 200-µl volume.
The following primary antibodies were used: mouse anti-BMI1 (Abcam89, Abcam), goat anti-MEL18 (ab5267, Abcam), rabbit anti-CBX8 (Bethyl Laboratories), rabbit anti-CBX7 (Abcam73, Abcamouseouse anti-p16INK4a (JC8), mouse anti-RING2 and mouse anti-HPH2 (both kindly provided by Haruhiko Koseki), rabbit anti-β-tubulin (H-235, Santa Cruz) and horse radish peroxidase (HRP -conjugated anti-FLAG M2 (Sigma).
The macrophage line Raw-Myd88GFP, TLR9 KO cells (both kindly provided by Dr. Hermann Wagner, Institute of Medical Microbiology, Immunology and Hygiene, Munich, Germany), the J774 macrophage line, peritoneal macrophages (collected from Balb/c mice by peritoneal lavage with PBS), and spleen-derived dendritic cells (SDCs) were used for studying the cellular uptake and trafficking of plasmid DNA.
Enzymatic hydrolysis was carried out using the commercial enzyme Cellic CTec2 or Cellic CTec3, both kindly provided by Novozymes investment Co.
Two cell-permeable, fluorescent substrates were used: red fluorescent tetramethylrhodamine-Star and the green fluorescent BG-505 (both kindly provided by Andreas Brecht, formerly Covalys Biosciences, Basel, Switzerland).
Oxygen bleached and unbleached birch pulp fibres (kindly provided by Eka Chemicals) both with and without hemicellulose removal (Table 2) were used for binding experiments with CBM-FITC.
