Fig. 2: The RNF121^M158R mutation reduces RNF121 protein levels, stability and localization to the cis-Golgi.

a Immunofluorescent staining using a: an anti-Myc-tag antibody to identify RNF121 co-expressed with the Myc tag; GM130 antibody as a cis-Golgi marker; Golgin-97 antibody as a trans-Golgi marker; and, PDI antibody as an endoplasmic reticulum (ER) marker, in a human MYCN amplified neuroblastoma cell line, Kelly. Scale bar: 100 μm. b Kelly cells transiently transfected with Myc-tagged wild-type RNF121 (RNF121^WT) or mutant RNF121^M158R for 48 h. Shown here are representative images of immunofluorescent staining using an anti-Myc-tag antibody for the RNF121 protein and a GM130 antibody for the cis-Golgi. Scale bar: 20 μm. c Immunoblot analysis of RNF121 expression in human MYCN amplified neuroblastoma cell line, SK-N-BE(2)-C, across the 4 days following transfection with either RNF121^WT or mutant RNF121^M158R. d Immunoblot analysis of RNF121 expression in human MYCN non-amplified neuroblastoma cell line, SH-SY5Y, following transfection with either RNF121^WT wild type or RNF121^M158R.