Fig 2: L1-polysome and L1 RNP complexes in Mael-/- testes.(A) Western blot analysis of L1 ORF1p and RPS6 in Mael-/- testicular extracts fractionated through 10–50% sucrose gradients in the presence of cycloheximide (CHX). Fractionated Mael+/- extract served as a negative control. The absorbance profile of the gradient is shown in red. (B) qRT-PCR analysis of L1 RNA (L1 ORF1 amplicon) and beta-actin mRNA (Actb) in Mael-/- testicular extracts fractionated through 10–50% sucrose gradients in the presence of CHX. RNA amounts per fraction are reported as percentages of the total RNA detected across the gradient; barplot shows mean percentage values + SEM of four biological replicates. (C) Western blot analysis of L1 ORF1p and RPS6 in Mael-/- testicular extracts fractionated through 10–50% sucrose gradients in the presence of EDTA (+ EDTA); the absorbance profile of the gradient is shown in blue. Upon addition of RNases A and T1 (+ EDTA + RNase) ribosomes are degraded (profile in green) and L1 RNPs disaggregated. (D) qRT-PCR analysis of L1 RNA (L1 ORF1 amplicon) and beta-actin mRNA (Actb) in Mael-/- testicular extracts fractionated through 10–50% sucrose gradients in the presence of EDTA. Barplot shows mean percentage values + SEM of three biological replicates. See also S2 Fig.