Ab52644

Eight paraffin blocks showing continuous histopathological transition from distant endometriosis, contiguous atypical endometriosis and ovarian clear cell carcinomas were selected for sectioning. Five of the blocks were genotyped as carrying the C/C genotype, three as carrying T/T genotype at rs11614913 in MIR196A2, and were utilized for this study. Immunofluorescence staining was performed to detect active cell nucleoli and ribosome biogenesis activity using 1:100 rabbit anti-nucleophosmin (anti-NPM; ab52644) and anti-nucleolin (anti-NCL; ab129200) monoclonal antibodies (Abcam PLC, Cambridge, MA). Immunostaining was independently scored by two pathologists, and specific nucleolus staining was scored as: negative (0), weakly positive (1+), moderately positive (2+) or strongly positive (3+). We used a combination of the percentage of positively stained cells and the intensity of nucleolus staining for statistical analysis. The H-score = ΣPi xi was calculated, where i is the intensity of the stained tumor cells (0 to 3+) and Pi is the percentage of the stained tumor cells for each intensity group (0 to 100%) as previously described [67 (link)]. For discordant cases, a third investigator was brought in to score and the final intensity score was determined by the majority scores.

Rabbit polyclonal antibodies (pAb) against Myc (R1208-1), GFP (SR48-02), FLAG (091 (link)2-1), and mouse mAbs against β-actin (M1210-2) and GST (M0807-1) were purchased from Huaan Biological Technology (Hangzhou, China). Rabbit pAbs against histone H3 (R1105-1) and β-tubulin (0807-2 (link)) were also purchased from Huaan Biological Technology. Mouse anti-Myc (05-419) and anti-FLAG (F1804) mAbs were obtained from Sigma-Aldrich (St. Louis, MO, USA). Anti-FLAG affinity resin (A2220) for immunoprecipitation was also purchased from Sigma-Aldrich. Rabbit mAb against NPM1 (ab52644) was purchased from Abcam (Cambridge, MA). Mouse mAbs to Cap, Rep, ORF3, and ORF4 of PCV2 were produced by our laboratory [24 (link),36–38 (link)]. NP-40 cell lysis buffer (50 mM Tris [pH 7.4], 150 mM NaCl, 1% NP-40) was purchased from Beyotime (P0013F; Shanghai, China). Horseradish peroxidase (HRP) or fluorescein isothiocyanate (FITC)-labeled goat anti-mouse and anti-rabbit IgG were purchased from KPL (Milford, MA, USA). Alexa Fluor 546-conjugated anti-mouse or anti-rabbit IgG were obtained from Invitrogen (USA).

Rabbit polyclonal antibodies (pAb) against Myc (R1208-1), GFP (SR48-02), Flag (0912-1), and mouse monoclonal antibodies (mAb) against β-actin (M1210-2) and GST (M0807-1) were purchased from Huaan Biological Technology (Hangzhou, China). Mouse anti-Myc (05-419) and anti-Flag (F1804) mAbs were obtained from Sigma-Aldrich (St. Louis, MO, USA). Anti-Flag affinity resin (A2220) for immunoprecipitation was also purchased from Sigma-Aldrich. Rabbit mAb against NPM1 (ab52644) was purchased from Abcam (Cambridge, MA). NP-40 cell lysis buffer (50 mM Tris [pH 7.4], 150 mM NaCl, 1% NP-40) was purchased from Beyotime (P0013F; Shanghai, China). Horseradish peroxidase (HRP)-labeled goat anti-mouse and anti-rabbit IgG were purchased from KPL (Milford, MA, USA).