Extended Data Fig. 8: The construction of RibosomeST knockout N2a cell line.
From: A male germ-cell-specific ribosome controls male fertility
a, Schematics of CRISPR-Cas9 and sgRNA against Rpl39l used in N2a cells. b, Sequences of the RibosomeST-/- N2a cell line generated by CRISPR-Cas9. c, Absorbance of ribosome profiles in RibosomeST-/- and wild-type N2a cell lines. d, Validation of TCP11 (P = 0.028), DCUN1D1 (P = 0.006) and DYNC1LI2 (P = 0.916) in RibosomeST-/- N2a cells by western blotting. n = 3. e, Absolute quantification of RPL39 and RPL39L protein expression levels in GC-1 ribosomes, and in RibosomeST-/- and wild-type N2a ribosomes by PRM (P = 0.004, n = 3). f, Quantification of Rpl39 mRNA in RibosomeST-/- and wild-type N2a cells (P = 0.034, n = 3). g, Western blotting of nascent peptides using anti-Puromycin in N2a cell lines following O-propargyl-puromycin assays, β-Tubulin was used as a control. n = 3. h, Quantification of Prm2, H1fnt, Ccin, and Dync1li2 in cell lines and testis. n = 3. For d, e, f and h, data are mean ± s.e.m. P values were determined using two-tailed Student’s t-tests. n values represent the number of independent experiments. Gel source data are provided in Supplementary Fig. 1.
