Antibodies and Plasmids for CSPP Research

Antibodies used in this study were as follows: anti-CSPP-L (Proteintech Europe, Manchester, United Kingdom; used for IF at 1 : 1000), anti-CSPP/CSPP-L (described in (Patzke et al, 2010 (link)); IF 1 : 200), anti-cytokeratin-8 (AbCam, Cambridge, UK; IF 1 : 500), anti-smooth muscle actin (Abcam; IF 1 : 100), anti-Cyclin A (Santa Cruz Biotechnology, Santa Crux, CA, USA; IF 1 : 1000), anti-γ-Tubulin (Sigma, St Louis, MO, USA; IF 1 : 500), and anti-Pericentrin (Abcam; IF 1 : 400). Secondary fluorochrome conjugated antibodies (Donkey anti-rabbit DyLight488; Donkey anti-mouse DyLight549; IF 1 : 1000) were purchased from Jackson Immuno Research (West Grove, PA, USA). shRNA and GFP co-expression plasmids used for CSPP1 knockdown were obtained from SA Biosciences (part of Qiagen N.V., Venlo, Netherlands): set KH18087G with the following target sequences: shRNA_01: 5′-gcacgaattcagcaggagtat-3′, shRNA_02: 5′-tccttcagttgacagcatcat-3′, shRNA_03: 5′-ggtgccaaagttgacttagat-3′, shRNA_04: 5′-ggaggtgaagatcgagaactt-3′, and shRNA_control: 5′-ggaatctcattcgatgcatac-3′. Tagged CSPP and CSPP-L full-length and truncation protein expression plasmids pCSPPmyc (derived from AJ583433) and pCSPP-Lmyc (derived from AM156947), pCSPP(498-876-eGFP) were described earlier (Patzke et al, 2006 (link)).