Enhanced chemiluminescence detection reagent

A Western blotting analysis was performed as previously reported [48 (link)]. The membranes were challenged with anti-POLR1B (Abcam, Cambridge, UK), anti-LC3B, anti-p62, anti-HA, anti-GAPDH and anti-β-actin (Cell Signaling Technology, Danvers, MA, USA). Proteins were visualized with enhanced chemiluminescence detection reagent according to the manufacturer’s instructions (Elabscience^®, Houston, TX, USA).

Immunoprecipitation assay was performed as previously reported^{48 (link)}. Briefly, whole cell lysate (1 mg) was incubated with 30 μl of protein A/G agarose beads coated with 5 μg of anti-uL3 (Primm, Milan, Italy) at 4 °C for 12 h. The beads were washed and boiled in the SDS sample buffer. The eluted proteins were loaded on 12% SDS-PAGE and detected by western blotting.
Western blotting analysis was performed as previously reported^{49 (link)}. The membranes were challenged with anti-E2F1 (Elabscience), anti-uL3 (Primm, Milan, Italy), anti-PARP-1, anti-GAPDH (Cell signaling), anti-B23, anti-p21, anti-α-tubulin, anti-β-actin, anti-nucleolin, anti-vinculin, anti-HA (Santa Cruz Biotechnology). Proteins were visualized with enhanced chemiluminescence detection reagent according to the manufacturer’s instructions (Elabscience®).

Protein extracts were prepared as previously described [30 (link)]. Western blotting analysis was performed as previously reported [31 (link)]. The membranes were challenged with anti-uL3, anti-eL8 (Primm, Milan, Italy), anti-uL5, anti-uL18 (Cell Signaling Technology, Danvers, MA, USA), anti-Bax, anti-p21, anti-CycB1, anti-α-tubulin, anti-α-actin, anti-GAPDH, and anti-Vinculin (Santa Cruz, Dallas, TX, USA). Proteins were visualized with enhanced chemiluminescence detection reagent according to the manufacturer’s instructions (Elabscience®, Houston, TX, USA).