Anti fibrillarin

The following primary antibodies were used: mouse monoclonal antibodies against LB1 (Zymed, San Francisco, CA, USA); LB2 (Abcam, Cambridge, MA, USA); LA/C (Millipore, Billerica, MA, USA); rabbit polyclonal antiserum against LB1 (Abcam); LA/C (Santa Cruz Biotechnology, Santa Cruz, CA, USA), trimethyl histone H3 (Lys27) (Millipore); γ-tubulin, β-tubulin and β-actin (Sigma, St. Louis, MO, USA); and BU1/75 (ICR1) rat monoclonal anti-BrdU antibody (Abcam). To analyze the localization of nuclear components, the following primary antibodies were used: anti-Nup153 (Abcam), anti-nuclear pore complex (mab 414; Covance, Princeton, NJ, USA), anti-sc-35 (Sigma), anti-LAP2β (BD Biosciences, San Diego, CA, USA), and anti-fibrillarin (Cytoskeleton Denver, CO, USA) mouse monoclonal antibodies, as well as antiactivated RNA Polymerase II monoclonal IgM (Covance).
For the Western blot analysis, horseradish-peroxidase-conjugated secondary antibodies (Bio-Rad Laboratories, Hercules, CA, USA) were used for detection. For the immunofluorescence analyses, Alexa fluorophore-conjugated anti-mouse, anti-rabbit, or anti-rat antibodies from Invitrogen were used.
Unless otherwise specified, the general reagents and chemicals were from Sigma, and the reagents for the cell cultures were from Invitrogen.

Drosophila Kc cells were maintained at 27 °C as logarithmically growing cultures in Schneider's medium (Sigma) + FBS (Gemini), and fixed and stained as previously described [66 (link)]. Antibodies used were anti-Fibrillarin (Cytoskeleton, catalog number AFB01; 1:200) and anti-H3K9me2 (Upstate, catalog number 07-442; 1:500).