Anti nucleolin

Equal amounts of protein from each sample were separated by 4–20% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to nitrocellulose membranes, and incubated with the following primary antibodies: anti-axin2 (Cell Signaling, #2151), anti-β-catenin (Cell Signaling, #8480), anti-lactate dehydrogenase A (Cell Signaling, #2012), anti- phosphoglycerate kinase 1 (Santa Cruz, sc-130335), anti-glyceraldehyde 3-phosphate dehydrogenase (Santa Cruz, sc-25778), anti-nucleophosmin (Abcam, ab10530), anti-nucleolin (Cell Signaling, 14574), anti-drebrin 1 (Abnova, H00001627-M03), anti-glycogen synthase kinase 3 α/β (Cell Signaling, #5676), and anti-gelsolin (Cell Signaling, #12953). Anti-β-actin (Sigma-Aldrich, A5441) was used as a loading control. After washing, the blots were labeled with horseradish peroxidase-conjugated secondary antibodies and visualized with the ECL reagent (GE Healthcare) according to the manufacturer’s protocol.

Primary antibodies used are as follows: ant-BANF1 (Abcam: ab88464, monoclonal, Sigma: SAB1404629, monoclonal), anti-HexaHis (Abcam ab1187), anti-FLAG M2 (Sigma, F3165), anti-Histone H3 (Cell Signaling, 9715), anti-nucleolin (Cell Signaling, 12247S) and anti-Emerin (Cell Signaling, 5430S). Fluorescent secondary antibodies used are: Donkey anti-Mouse 800 nm (LiCor; IRDye 800CW 926-32212), Donkey anti-Rabbit (LiCor; IRDye 680LT 926-28023) and Alexa Fluor 488 and 594 (Molecular Probes).

For western blot assay, cell lysates were sonicated and boiled at 95°C for 5 min as previously described (Ho et al. 2015 (link)). All membranes were incubated with the following primary antibodies: anti-LRRK2 [N241A/34] (75-253, Neuromab), anti-LRRK2 pS935 (ab133450, Abcam), anti-LRRK2 pS1292 (ab230181, Abcam), anti-c-myc [9E10] (sc-40,Santa Cruz), anti-nucleolin (#14574, Cell Signaling Technology [CST]), anti-hsp70 (sc-66048, Santa Cruz), anti-moesin (sc-13122, Santa Cruz), anti-moesin pT558 (ab61109, Abcam), anti-PARP (9542S, CST), anti-α-tubulin (T9026, Sigma-Aldrich), and anti-β-actin (sc-47778, Santa Cruz). After primary antibody reaction, membranes were incubated in goat anti-rabbit or anti-mouse IgG with horseradish peroxidase (Jackson Immunoresearch). Intensities of the bands were detected by the Multi-Gauge V 3.0 program (Fuji photo Film).