Fig. 2: RSL24D1 is associated with pre-60S subunits in mouse ESCs.
a Comparison of the mouse RSL24D1 predicted structure (amino acids 1–135, red color) with the yeast Rlp24 structure (amino acids 1–149, gray color, left panel) and the human RSL24D1 structure (amino acids 1–163, blue color, right panel)33,34. The mouse RSL24D1 protein structure was predicted from the 6N8J and 6LSS PDB structures using the swiss-model structure assessment tool69. b Representative images of naive CGR8 cells stained with Hoechst and with anti-FBL and anti-RSL24D1 antibodies (20× objective) (n = 3). The scale bar represents 10 μm. c Representative immunoblots of ESCFBS nucleo-cytoplasmic fractions before (total) and after sucrose cushion purifications of nuclear pre-ribosomes (pre-rib.) and cytoplasmic ribosomes (ribosomes) (n = 3). HISTONE 3 and GAPDH are shown as specific nuclear and cytoplasmic proteins, respectively. d Polysome profiling of CGR8 ESCFBS cytoplasmic extracts. Ribosome-free fractions (free mRNPs), 40S, 60S, 80S monosomes and polysomes are detected by UV-absorbance and indicated on the absorbance curve (upper panel). Representative immunoblots of gradient fractions (middle panel) (n = 3). GAPDH is used as a control of the free mRNPs. Total RNAs were extracted from collected fractions, analyzed on non-denaturing agarose gels, and revealed with ethidium bromide. The positions of the 28S, 18S, 5.8S, 5S rRNAs, and tRNAs are indicated (lower panel).
