RPL39 Antibody - #DF3711

Fig. 1. Expression of Rpl39 and Rpl39l paralogs in NIH3T3 cells. (A) RT-PCR showing that both Rpl39 and Rpl39l are expressed in NIH3T3 cells. Gapdh was used as the internal control. (B) Standard curves of transcript levels of Rpl39 and Rpl39l. Series dilutions of known quantities of Rpl39 and Rpl39l cDNAs were used in quantitative RT-PCR with fluorescently tagged gene specific primers. CT values were plotted against calculated copy numbers of Rpl39 and Rpl39l transcripts. (C) Determination of copy numbers of Rpl39 and Rpl39l transcripts in NIH3T3 cells. Total RNA extracted from NIH3T3 cells were used in quantitative RT-PCR with fluorescently tagged gene specific primers. Copy numbers of transcripts were calculated according to the stand curve shown in (B). Shown are averages of four independent experiments. 100 ng of total RNA was used in each experiment. (D) Gene targeting of Rpl39 and Rpl39l in NIH3T3 cells using CRISPR/Cas9 method. Schematic drawings show genomic organizations of the two genes. sgRNA targeted sequences are underlined. The resulting deletion mutations are shown below the wild type sequences, with the sequencing data on the right. The double mutations of Rpl39 and Rpl39l is generated in the Rpl39 -/- cells. Positions of gene-specific primers for genotyping are indicated by red arrows. Red arrowheads show the positions of the single nucleotide deletion. (E) Amino acid sequences of RPL39 and RPL39L. Both proteins are 51-aa long, with only three amino acids are different (blue letters). The mutants generated from gene targeting would result in aberrant peptides or early stop, as shown in red. The peptide used by Affinity for generating the antibody against RPL39 is indicated by black line. (F) Western blotting of NIH3T3 cells. Cell lysates from wild type and mutant cells were subjected to SDS-PAGE and immuno-blotted with Affinity anti-RPL39 antibody. An increase of RPL39 in Rpl39l -/- cells can be seen.