Alexa flour 488 conjugated goat anti mouse

The following antibodies were used: MV Phosphoprotein (P) (ARGENE); MV matrix (M) (Abcam); CDK6 (Santa Cruz Biotechnology). CDK4 (Santa Cruz Biotechnology) is a functional homologue of CDK6 that is not a target for miRNA-124. Antibodies against it serve as a negative control in Western blots. CopGFP (Enzo Life Sciences & Axxora); GAPDH (MILLIPORE); β-actin (SIGMA-ALDRICH); anti-mouse and anti-rabbit IgG peroxidase Jackson ImmunoResearch Alexa flour®-488 conjugated goat anti-mouse and Alexa flour®-647 conjugated goat anti-rabbit (Molecular Probes Inc.). Fluorescence tags included Alexa Fluor 647-Annexin V, 7-AAD (BioLegend); DAPI (Sigma) and TO-PRO (ThermoFisher).

HL-derived L428 and KMH2 cells were grown in suspension in RPMI 1640 supplemented with 100 U/ml penicillin, 0.1 mg/ml streptomycin, 2 mM L-glutamine and 10% fetal calf serum (FCS), in a 5% CO2 humidified atmosphere at 37 °C. For the induction of senescence cells were treated with 50 μM H₂O₂ for 2 h in RPMI without FCS, washed with complete medium and grown for 4–7 days.
Antibodies used included: anti-p21 ^Cip1 (sc-397), anti-p16 ^INK4a (sc-1207), anti-p53 (sc-126) and anti-p65 (sc-8008) and phospho-NF-κB p65 (Ser 536) (sc-33020- R) which were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), anti-γH2AX phosphor -S139 (ab18311, Abcam, Cambridge, MA, USA), anti-trimethyl Histone H3 (Lys4/Lys9) (Millipore 07-992). Anti CD15 LeuM1 and anti-CD30 BerH2 (Dako, Copenhagen, Denmark), Ki-67, (ab15580, Abcam). For florescence detection the following secondary antibodies were used: donkey anti-rabbit IgG (DyLight 549, 711-505-152, Jackson ImmunoResearch Laboratories, West Grove, PA, USA), Alexa flour-488 conjugated goat anti-mouse (Molecular Probes Inc., Eugene, OR, USA).