Article Title: Inhibition of Poly-ADP-Ribosylation Fails to Increase Axonal Regeneration or Improve Functional Recovery after Adult Mammalian CNS Injury
Figure Lengend Snippet: Selective suppression of Parp1 improves cortical neuron axonal regeneration. A , Representative images of regenerated cortical neurons. Neurons were transduced with shRNA lentiviral particles on culture d 3 and scraped on d 8. Scraped neurons were fixed and stained at d 11. The microphotographs illustrate βIII tubulin in axons (green), phalloidin of F-actin in growth cones (phalloidin, red), and cell nuclei (DAPI, blue). Note that no neuronal cell bodies (DAPI) migrate into the scrape zone. Selectively after Parp1 knockdown, a greater number of regenerating axons and growth cones are visible in the scrape zone. Scale bars, 200 µm. B , Quantification of axon regeneration. Data are mean ± SE. shNC, n = 109; sh Parp1 , n = 49 (each of 5 lentiviral species has 10 independent wells, with 1 cell culture lost); sh Parp2 , n = 10 (each of 5 lentiviral species has 2 independent wells); sh Parp3 , n = 10 (each of 5 lentiviral species has 2 independent wells); sh Parp9 , n = 10 (each of 5 lentiviral species has 2 independent wells); sh Parp12 , n = 10 (each of 5 lentiviral species has 2 independent wells); and sh Parp16 ; n = 16 (each of 4 lentiviral species has 4 independent wells). Each condition was compared to NC control by one-way ANOVA followed by Dunnett test. **, P
Article Snippet: On day in vitro 3 (DIV3), 1 × 105 TU of lentiviral particles targeting mammalian nontargeting (NC) short hairpin RNA (shRNA) control (cat. #SHC002V), Parp1 shRNA (NMID: NM_007415, clone ID: TRCN0000071208, TRCN0000071209, TRCN0000071210, TRCN0000071211, TRCN0000071212; Sigma-Aldrich), Parp2 shRNA (NMID: NM_009632, clone ID: TRCN0000071213, TRCN0000071214, TRCN0000071215, TRCN0000071216, TRCN0000071217; Sigma-Aldrich), Parp3 shRNA (NMID: NM_145619, clone ID: TRCN0000093894, TRCN0000093895, TRCN0000093896, TRCN0000093897, TRCN0000093898; Sigma-Aldrich), Parp9 shRNA (NMID: NM_030253, clone ID: TRCN0000174399, TRCN0000174697, TRCN0000173214, TRCN0000176202, TRCN0000175373; Sigma-Aldrich), Parp12 shRNA (NMID: NM_172893, clone ID: TRCN0000174741, TRCN0000174854, TRCN0000175447, TRCN0000175542, TRCN0000175901; Sigma-Aldrich), or Parp16 shRNA (NMID: NM_177460, clone ID: TRCN0000200923, TRCN0000190330, TRCN0000201597, TRCN0000190801; Sigma-Aldrich) were added to primary cortical neurons.
Techniques: Transduction, shRNA, Staining, Cell Culture