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. 2022 Oct 26;23(21):12909. doi: 10.3390/ijms232112909

Figure 3.

Figure 3

Chronic 3HB exposure interferes with insulin signaling in cardiomyocytes. ARCMs were cultured for 24 h in either low palmitate medium (LP; control condition), high palmitate medium (HP), LP with 3 mM 3HB (3HB), or LP with 3HB in the presence of 4*KLR (3HB/4*KLR). (A,B) After 24 h, cells were short-term (30 min) incubated without/with 100 nM insulin. Thereafter, (A) Ser473 phosphorylation of Akt, (B) Ser2447 phosphorylation of mTOR, (C) Thr 642 phosphorylation of AS160, and (D) Ser235/236 phosphorylation of S6 were assessed by Western blotting and quantified. For this, normalization was performed against loading controls. Loading control for the degree of phosphorylation of Akt and AS160 is caveolin-3. Loading control for mTOR and S6 phosphorylation is GAPDHs. Representative blots and corresponding loading controls are displayed (n = 8). Bar values are means ± SEM. * p < 0.05. ns: not significant. The dots represent the measurements that are performed under ‘basal’ condition. The triangles represent the measurements that are performed after insulin stimulation.