10.6084/m9.figshare.c.3629300_D4.v1 Kristine Kines Kristine Kines Mark Sokolowski Mark Sokolowski Dawn deHaro Dawn deHaro Claiborne Christian Claiborne Christian Melody Baddoo Melody Baddoo Madison Smither Madison Smither Victoria Belancio Victoria Belancio Additional file 7: Figure S4. of The endonuclease domain of the LINE-1 ORF2 protein can tolerate multiple mutations Springer Nature 2016 LINE-1 L1 ORF2 Endonuclease Mutation Retrotransposition Phosphorylation 2016-04-19 05:00:00 Figure https://springernature.figshare.com/articles/figure/Additional_file_7_Figure_S4_of_The_endonuclease_domain_of_the_LINE-1_ORF2_protein_can_tolerate_multiple_mutations/4418576 Western blot analysis of ORF2 proteins containing mutations in putative phosphorylation sites. Top panel: Representative western blot analysis of total cell lysates harvested from HeLa cells transfected with the indicated ORF2 putative phosphorylation mutant constructs. ORF2 is the functional protein and ORF2 RT- is a non-functional protein containing a mutation in the reverse transcriptase (D702A) domain. Control lanes indicate cells transfected with an empty vector. Lysates were probed with anti-γH2AX antibodies to detect the phosphorylation of histone H2AX in response to DNA damage, top; and anti-GAPDH to serve as a loading control, bottom. Bottom panel: Western blot quantitation. For each sample, the signal detected for γH2AX was normalized to the signal detected for GAPDH. These relative numbers were expressed as a proportion of the relative number detected from the functional ORF2p. (PNG 263 kb)