A Mouse brain kDa Control Atg7 CKO 250- 50- 50- 50- 37- E MEF with starvation Atg7 WT Atg7 KO kDa 0 3 6 9 0 3 6 9h 250- 50- 50- 15- 37- F Relative protein level +AKAP11 + Rla 1.5* Rila 1.0 0.5- 0.0+ 0 3 6 Atg7 WT -AKAP11 -Rila -Rla -p62 -Actin gh Relative protein level 1.5 1.0 0.5- B Control Atg7 CKO 8₁ 0.0 Relative Protein level -AKAP11 -Rila -Rla -LC31 -LC311 -Actin + ल 2 AKAP11 +AKAP11 +Rla Rila J Ria ns Rila с kDa 250- 50- 50- 50- 37- G 50- 50- HEK293T with starvation Atg7 WT Atg7 KO kDa 0 3 6 9 0 3 6 9h 250- 15- 37- H Relative protein level Mouse brain Control Atg14 CKO 1.0 +Rla 1.5 Rila 0.5- +AKAP11 0.0+ # # 6 9h -AKAP11 -Rila -Ria -p62 -Actin 3 Atg7 WT Relative protein level 1.0 Relative Protein level 0.5- 8- 0.0+ 9.00 99 -AKAP11 -Rila -Ria LLC31 FLC311 -Actin 2 1.5 Rila *² Control Atg14 CKO AKAP11 AKAP11 - Rla J Rla 3 6 gh Atg7 KO Fig. 1. AKAP11 and Rla are degraded by autophagy. (A) Brain lysates of Atg7FF (Control) and Atg7fF-SynCre (CKO) were subjected to immunoblotting analysis with the indicated antibodies. (B) Quantification of protein levels in A. Unpaired Student's t tests were used, and values are presented as mean + SEM (n = 3 mice/genotype). **P < 0.01; ***P < 0.001; ns, not significant. (C) Brain lysates of Atg14F (Control) and Atg14FF-SynCre (cKO) were subjected to immunoblotting analysis with the indicated antibodies. (D) Quantification of protein levels in C. Unpaired Student's t tests were used, and values are pre- sented as mean ± SEM (n = 4 mice/genotype). **P<0.01; ***P<0.001; ns, not significant. (E) Atg7 WT and Atg7 KO MEF cells were nutrient starved by using EBSS for the indicated times, and cells were assayed by immunoblotting analysis with the indicated antibodies. (F) Quantification of the protein levels from E was obtained by normalizing the level of proteins to Actin, respectively, and further to the respective control. One-way ANOVA was used in each genotype, and values are presented as mean ± SEM (n = 3). *P<0.05; **P<0.01. (G) Atg7 WT and Atg7 KO HEK293T cells were nutrient starved by using EBSS for the indicated times and were then assayed by immunoblotting analysis with the indicated antibodies. (H) Quantification of the protein levels from G was obtained by normalizing the level of proteins to Actin, respectively, and further to the respective control. One-way ANOVA was used in each genotype, and values are presented as mean ± SEM (n = 4). "P<0.05; **P < 0.01; ***P < 0.001. 3 6 9h Atg7 KO ns Rila CELL BIOLOGY
Autophagy Is Required for PKA Activation and Cell Viability upon Glucose
Starvation. The functional relationship between PKA and autophagy
is largely unclear. Because PKA controls cell
metabolism with glucose limitation. We found that in addition to
activating autophagy, glucose starvation also induced the phosphorylation of cAMP response element-binding (p-CREB), a wellknown PKA substrate, indicative of elevated PKA activity in both
HEK293T and HCT116 cells. In contrast, disruption of autophagy
by deletion of essential autophagy gene Atg7 or Atg14 abolished the
increase of p-CREB levels in the above cells (SI Appendix, Fig.
S1 A, B, D, and E), suggesting that inactivation of autophagy suppresses PKA activity in response to energy shortage. Moreover, loss
of Atg7 or Atg14 accelerated cell death in response to glucose
starvation (SI Appendix, Fig. S1 C and F). Taken together, our results show that autophagy is required for PKA activation and important for cell viability under metabolic stress.
Fig. 1. AKAP11 and RIα are degraded by autophagy. (A) Brain lysates of Atg7F/F (Control) and Atg7F/F-SynCre (cKO) were subjected to immunoblotting analysis with the indicated antibodies. (B) Quantification of protein levels in A. Unpaired Student's t tests were used, and values are presented as mean ± SEM (n = 3 mice/genotype). **P < 0.01; ***P < 0.001; ns, not significant. (C) Brain lysates of Atg14F/F (Control) and Atg14F/F-SynCre (cKO) were subjected to immunoblotting analysis with the indicated antibodies. (D) Quantification of protein levels in C. Unpaired Student's t tests were used, and values are presented as mean ± SEM (n = 4 mice/genotype). **P < 0.01; ***P < 0.001; ns, not significant. (E) Atg7 WT and Atg7 KO MEF cells were nutrient starved by using EBSS for the indicated times, and cells were assayed by immunoblotting analysis with the indicated antibodies. (F) Quantification of the protein levels from E was obtained by normalizing the level of proteins to Actin, respectively, and further to the respective control. One-way ANOVA was used in each genotype, and values are presented as mean ± SEM (n = 3). *P < 0.05; **P < 0.01. (G) Atg7 WT and Atg7 KO HEK293T cells were nutrient starved by using EBSS for the indicated times and were then assayed by immunoblotting analysis with the indicated antibodies. (H) Quantification of the protein levels from G was obtained by normalizing the level of proteins to Actin, respectively, and further to the respective control. One-way ANOVA was used in each genotype, and values are presented as mean ± SEM (n = 4). # P < 0.05; **P < 0.01; ***P < 0.001.
can you please explain figure A-H I dont understand what it represents
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