α β actin ac 15

The following primary antibodies were utilized: mouse (ms) α α-synuclein (α-syn) (BD/610787), rabbit (rb) α GGA1 (H215, Santa Cruz/sc-30102), ms α GGA2 (BD/612612), ms α GGA3 (BD/612310), ms α c-myc (9E10, Sigma-Aldrich/M4439), ms α β-actin (AC-15, Sigma-Aldrich/A5441), rb α FLAG (Sigma-Aldrich/F7425), rb α tyrosine hydroxylase (TH) and ms α Flotillin-1 (BD/610820). Secondary horseradish peroxidase (HRP)-coupled antibodies were obtained from Molecular Probes/Invitrogen.
Fusion constructs of α-syn-hGLuc1 (S1), α-syn-hGLuc2 (S2), [87 (link), 88 (link)] as well as VPS4_WT and VPS4_DN constructs (kind gift of Prof. Dr. Nobuyuki Tanaka) [32 (link), 89 (link)] have been previously described. GGA2-myc and GGA3-myc plasmids [79 (link)] were generated by cloning into pcDNA3.1(−) Myc/His A (Invitrogen) as previously described for GGA1-myc [57 (link)]. GGA3-mRFP was generated by cutting GGA3 cDNA out of the GGA3-myc construct using NheI and EcoRI cleavage site and subcloning into NheI and EcoRI sites of mRFP-N1 (Roger Tsien).

Monoclonal antibodies used for immunofluorescence and Western blot analyses were: α-IE1 63–27 [81 (link)], α-UL44 BS510 (kindly provided by B. Plachter, Mainz, Germany), α-MCP 28–4 [82 (link)], α-FLAG M2 (Sigma-Aldrich, Deisenhofen, Germany), α-Myc 9E10, α-β-actin AC-15 (Sigma-Aldrich, Deisenhofen, Germany), α-FEN1 B-4 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), α-phospho-Histone H2A.X (Ser139) (20E3) (Cell signaling technology, Danvers, MA, USA), α-E2F1 (KH95) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and α-BrdU MoBu-1 (Thermo Fisher Scientific, Waltham, MA, USA). The polyclonal antibodies α-phospho FEN1 (Ser187) (Thermo Fisher Scientific, Waltham, MA, USA) and α-p-Histone H2A.X (Ser 139) sc-101696 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and were used for Western blot analyses. Secondary antibodies used for immunofluorescence and Western blot analyses were: Alexa Fluor 488-/555-/647-conjugated secondary antibodies for indirect immunofluorescence experiments were purchased from Molecular Probes (Karlsruhe, Germany), horseradish peroxidase-conjugated anti-mouse/-rabbit secondary antibodies for Western blot analyses were obtained from Dianova (Hamburg, Germany).

Monoclonal antibodies used for immunofluorescence and Western blot analyses were: α-IE1 CH443 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), α-UL112/113 M23, α-UL44 BS510 (kindly provided by B. Plachter, Mainz, Germany), α-UL69 69–66, α-FLAG M2 (Sigma-Aldrich, Deisenhofen, Germany), α-Myc 9E10, α-β-actin AC-15 (Sigma-Aldrich), α-PML PG-M3 (Santa Cruz). Polyclonal antibodies used for immunofluorescence and Western blot analyses were: α-rhesus IE1 (a kind gift from M. Mach, Erlangen, Germany), α-PML #4 (a kind gift from P. Hemmerich, Jena, Germany), α-PML H238 (Santa Cruz), α-PML A301–167A (Bethyl Laboratories, Montgomery, TX, USA), α-PML A301–168A (Bethyl Laboratories), α-Sp100 #2 (a kind gift from P. Hemmerich, Jena, Germany), α-Sp100 GH3 (kindly provided by H. Will, Hamburg, Germany), α-hDaxx C-20 (Santa Cruz), α-ATRX H-300 (Santa Cruz). Secondary antibodies used for immunofluorescence and Western blot analyses were: Alexa Fluor 488-/555-/647-conjugated secondary antibodies for indirect immunofluorescence experiments were purchased from Molecular Probes (Karlsruhe, Germany), horseradish peroxidase-conjugated anti-mouse/-rabbit secondary antibodies for Western blot analyses were obtained from Dianova (Hamburg, Germany).