Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2015 Jun 1.

Published in final edited form as: Neurobiol Aging. 2013 Dec 14;35(6):1431–1441. doi: 10.1016/j.neurobiolaging.2013.12.005

Fig. 5 — Upregulation of ER stress markers with sleep deprivation (SD). (A) Densitometric quantification of BiP expression in young undisturbed (YUD), young sleep deprived (YSD), young PBA (YPBA) and young PBA sleep deprived (YPBASD) wCS10 fly heads (n=4 trials, 10 heads/pool). 6 h of SD increased BiP levels in young flies in comparison to their non-SD counterparts (p=0.007). Young flies treated with PBA responded similarly to young untreated flies when sleep deprived (p<0.05). (B) Representative western blots. 15µg of protein were loaded per well from undisturbed and sleep deprived flies. (C) Quantification of BiP expression in aged undisturbed (AUD); aged sleep deprived (ASD), aged PBA (APBA) and aged PBA sleep deprived (APBASD) fly heads. (D) Representative western blot shows 15µg of protein loaded per well from undisturbed and SD flies. BiP expression increased in aged sleep deprived flies (ASD) versus aged undisturbed flies (AUD) (p<0.05) (n=4 trials, 10 heads/pool). Aged flies treated with PBA showed no significant induction of BiP after sleep deprivation. All gels were stripped and reprobed with β-actin as the loading control. (E-F) ER stress triggers Drosophila XBP1 mRNA splicing, which is characterized by the elimination of a 23 base pair intron from Drosophila XBP1 mRNA. PBA reduced both the unspliced and spliced variant of XBP1 in the YPBASD and APBA flies. In the APBASD flies XBP1 splicing is decreased. Mean +S.E.M. shown for XBP1 *P<0.05, **P<0.01, #P<0.001 (G) Densitometric quantification of phosphoeIF2α levels in fly heads (n=4 trials, 10 heads/pool). 6 h of sleep deprivation (SD) increased peIF2α expression in YSD flies in comparison to their undisturbed counterparts (YUD) (p=0.032). Aged flies (AUD) have increased basal levels of p-eIF2α that are significantly higher than the YUD flies (p=0.006). There was no significant difference between the AUD and aged ASD flies; however, p-eIF2α levels are markedly reduced in aged SD flies with acute application of PBA (ASDPBA) (p<0.05). Mean +S.E.M. shown, *P<0.05, **P<0.01. (H) Representative Western blots of p-eIF2α shows 15µg of protein loaded per well from undisturbed and SD flies. All gels were stripped and reprobed with β-actin as the loading control.

Fig. 5 — Upregulation of ER stress markers with sleep deprivation (SD). (A) Densitometric quantification of BiP expression in young undisturbed (YUD), young sleep deprived (YSD), young PBA (YPBA) and young PBA sleep deprived (YPBASD) wCS10 fly heads (n=4 trials, 10 heads/pool). 6 h of SD increased BiP levels in young flies in comparison to their non-SD counterparts (p=0.007). Young flies treated with PBA responded similarly to young untreated flies when sleep deprived (p<0.05). (B) Representative western blots. 15µg of protein were loaded per well from undisturbed and sleep deprived flies. (C) Quantification of BiP expression in aged undisturbed (AUD); aged sleep deprived (ASD), aged PBA (APBA) and aged PBA sleep deprived (APBASD) fly heads. (D) Representative western blot shows 15µg of protein loaded per well from undisturbed and SD flies. BiP expression increased in aged sleep deprived flies (ASD) versus aged undisturbed flies (AUD) (p<0.05) (n=4 trials, 10 heads/pool). Aged flies treated with PBA showed no significant induction of BiP after sleep deprivation. All gels were stripped and reprobed with β-actin as the loading control. (E-F) ER stress triggers Drosophila XBP1 mRNA splicing, which is characterized by the elimination of a 23 base pair intron from Drosophila XBP1 mRNA. PBA reduced both the unspliced and spliced variant of XBP1 in the YPBASD and APBA flies. In the APBASD flies XBP1 splicing is decreased. Mean +S.E.M. shown for XBP1 *P<0.05, **P<0.01, #P<0.001 (G) Densitometric quantification of phosphoeIF2α levels in fly heads (n=4 trials, 10 heads/pool). 6 h of sleep deprivation (SD) increased peIF2α expression in YSD flies in comparison to their undisturbed counterparts (YUD) (p=0.032). Aged flies (AUD) have increased basal levels of p-eIF2α that are significantly higher than the YUD flies (p=0.006). There was no significant difference between the AUD and aged ASD flies; however, p-eIF2α levels are markedly reduced in aged SD flies with acute application of PBA (ASDPBA) (p<0.05). Mean +S.E.M. shown, *P<0.05, **P<0.01. (H) Representative Western blots of p-eIF2α shows 15µg of protein loaded per well from undisturbed and SD flies. All gels were stripped and reprobed with β-actin as the loading control.