Journal: Pediatric research

Article Title: Preterm birth alters the feeding-induced activation of Akt signaling in the muscle of neonatal piglets.

doi: 10.1038/s41390-022-02382-4

Figure Lengend Snippet: Fig. 2 Preterm birth attenuates the phosphorylation of Akt1 and Akt2, but not Akt3. The phosphorylation of individual Akt isoform was analyzed by immunoprecipitation followed by western blot. a–c Phosphorylated Akt1 at Ser472, Akt2 at Ser473, and Akt3 at Ser474. Phosphorylation values were corrected by their protein abundance in the immunoprecipitates. For Akt1 and Akt2, there was a significant interaction (P < 0.01) between GAB and STATE. Representative blots are shown. Data are expressed as mean ± SEM; n = 21–25/group (*P < 0.05 and **P < 0.01). AU arbitrary units.

Article Snippet: The primary antibodies that were used were: phospho-pan-/Akt (Ser472/473/474) (1:1000; #ab192623; Abcam, Cambridge, MA); pan-Akt, Akt1, Akt2, Akt3 (Rabbit mAb #4685, Rabbit mAb #75692, Rabbit mAb #3063, and Mouse mAb #8018), FAK (total, #3285 and Tyr397, #8556), mTOR (total, #2972), PTEN (total, #9552 and Ser308, #9551), PP2A (total, #2260) and PDK1 (total, #3062 and Ser241, #3438) (1:1000; Cell Signaling Technology); PP2A (Tyr307, # AF3989-SP; 1:1000; R&D System, Minneapolis, MN); Vinculin (#66305-1-lg; 1:4000); PHLPP (#676640-1-lg), Ubl4A (#14253-1-AP), IP6K1 (#12057-2-AP), Hsp90 (#13171-1-AP) and GAPDH (#60004-1-lg) (1:1000; Proteintech, Rosemont, IL); phospho-mTOR (Ser2481, #651701) (1:1000; BioLegend).

Techniques: Phospho-proteomics, Immunoprecipitation, Western Blot, Quantitative Proteomics