Rabbit anti sfpq

Rat anti-HBoV1 NS1C antibody was produced previously (108 (link)). The following primary antibodies were purchased: mouse anti-Flag (number 200-301-B13) from Rockland (Limerick, PA); rabbit anti-Strep-tag II (number ab183907) from Abcam (Waltham, MA); mouse anti-β-actin (number A5441) from MilliporeSigma; mouse anti-Ku70 (number SC-17789) and mouse anti-GST (number SC-138) from Santa Cruz (Dallas, TX); rabbit anti-Ku70 (number A7330), rabbit anti-HSPA8 (number A14001), rabbit anti-GTF2F1 (number A2489), rabbit anti-ORC3 (number A15415), rabbit anti-DNA-PKcs (number A1419), rabbit anti-DHX9 (number A4563), rabbit anti-SNW1 (number A14580), rabbit anti-PABPC1 (number A14872), rabbit anti-DDX17 (number A17078), rabbit anti-RPA1 (number A0990), and rabbit anti-MCM3 (number A1060) from Abclonal (Woburn, MA); rabbit anti-SFPQ (number A301-321A-T) from Bethyl (Montgomery, TX); rabbit anti-HSPA1b (number A59434-020) from EpiGentek (Farmingdale, NY); rat anti-HSPA8 (number NBP1-97868) from Novus Bio (Centennial, CO); anti-BrdU (clone B44) from BD Biosciences (San Jose, CA); and anti-GAPDH (number 2118S) from Cell Signaling (Danvers, MA).

CFBE cells were fixed and stained as previously described in Kumar et al.^{76 (link)}, with minor modification. CFBE cells were seeded onto a coverglass, precoated with poly-d-lysine hydrobromide (50 μg/ml) (Sigma, P7886). The cells were washed twice with phosphate-buffered saline (PBS) and then fixed for 15 min with 4% paraformaldehyde at room temperature. Fixative solution was aspirated and cells were washed three time with 1 × PBS for 5 min. Cells were then permeabilized with 0.2% triton X-100 for 10 min at room temp. Permeabilized cells were again washed with sterile 1× PBS, for 5 min and then blocked with blocking buffer (1% BSA + 0.3% triton X-100 in 1× PBS). Primary antibodies (mouse anti-CFTR, 1:500, R&D Systems and Rabbit anti- SFPQ, 1:750, Bethyl Laboratories), diluted in blocking buffer, were added to the cells and incubated overnight at 4 °C. Next day cells were washed and incubated for 2 h at room temperature in appropriate secondary antibodies. After washing, the cells were then incubated in DAPI for 15 min and the cover glass was mounted with Fluoromount-G (Invitrogen, 00-4958-02). Samples were imaged using a Zeiss 700 confocal microscope and an average of 300 cells in 10 different fields was analyzed.