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. 2019 Dec 3;77(18):3657–3670. doi: 10.1007/s00018-019-03392-y

Fig. 6.

Fig. 6

a Eclosion experiments show that coexpression of Orb2A and Orb2B can rescue the lethality associated with pan-neuronal expression of HttQ138 using Elav Gal4. In comparison to only HttQ138 (n = 608) which showed 4.8% eclosion, coexpression with CD8GFP (n = 26) showed 3.8% eclosion, while coexpression with Orb2A (n = 131) and Orb2B (n = 187) showed 60% and 51% eclosion respectively. b Eclosion experiments performed with a lower expressing Elav Gal4 line show that in the presence of Orb2RNAi coexpression there is decreased survival of HttQ138 expressing animals. For nonpathogenic HttQ15 coexpression with Luciferase (Luc) RNAi (n = 35) or Orb2 RNAi (n = 35) did not show any difference. HttQ138 coexpression with LucRNAi showed 77.8% survival (n = 185) and with Orb2 RNAi showed 50% survival (n = 165). c HttQ138 aggregate numbers in motor neurons in larvae expressing HttQ138 alone (left panel) or together with Orb2A and Orb2B (right panels) do not show any significant difference. d Quantitation of aggregate numbers from HttQ138 alone (n = 3, 11 axons) and HttQ138 with Orb2A (n = 3, 20 axons) or Orb2B from (n = 3, 18 axons) motor neurons do not show any significant difference. Error bars represent SEM and significance tested using unpaired one-tailed t test. e Representative SDD-AGE immunoblot showing no significant difference between the Htt oligomers in HttQ138 and HttQ138 with Orb2A and Orb2B. f Quantitation of Polysome/80S ratio from HttQ138 and HttQ138 + Orb2A GFP and HttQ138 + Orb2B GFP expressing cells shows that coexpression of Orb2A and Orb2B rescues the reduced polysome/80S ratio associated with HttQ138 cells. Data are from n = 3 experiments and represented as a relative fold change for others compared to HttQ15. Error bars represent SEM and significance is tested using unpaired one-tailed t test. g Quantitation of Puromycin incorporation from HttQ138 and HttQ138 + Orb2A GFP and HttQ138 + Orb2B GFP expressing cells shows significantly reduced Puromycin incorporation in HttQ138 cells and rescue with coexpression of Orb2A and Orb2B. Data are from n = 3 experiments and is represented as a relative fold change for others compared to HttQ15. Error bars represent SEM and significance is tested using unpaired one-tailed t test