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. 2021 Jun 3;22(11):6034. doi: 10.3390/ijms22116034

Figure 3.

Figure 3

mTORC1 inhibition in the embryonic cerebral cortex downregulates cholesterol biosynthesis genes in vivo. Pregnant mice received either a single injection according to E16.5 of the offspring or were injected on three consecutive days (E14.5-E16.5) once per day with DMSO or rapamycin at a dose of 1 mg/kg. Subsequently, total RNA and protein were extracted from the cerebral cortex and RT-qPCR and Western blot experiments were performed. (A) Inhibition of mTORC1 activity was confirmed by Western blot analysis of the mTORC1 downstream effector pS6 (compared to S6) 24 h after a single rapamycin injection. (B,D) RT-qPCR to detect RNA expression of the four genes Ldlr, Osc, Mvd, and Dhcr7 after one day (B) or three days (D) of i. p. injection of rapamycin compared to DMSO; mRNA expression was normalized to Gapdh. (C,E) Immunoblot analysis to detect Ldlr, Osc, Mvd, and Dhcr7 protein expression after one day (C) or three days (E) of i. p. injection of rapamycin compared to DMSO and normalized to Gapdh. Data represent the average of six biological replicates. For statistical analyses, Student’s t-test was used. * p < 0.05; ** p < 0.01; *** p < 0.001.