Fig. 5

FSS induces GOT-dependent amino acid metabolism in trophoblasts cells were cultured either under static, or flow conditions for 24 h. qPCR analysis of GOT1 (a). Representative Western Blot (b) and band densitometry (c) showed increasing GOT1 levels in BeWo cells exposed to shear stress. GOT2 mRNA (d) and protein levels (e and f) were upregulated in cells cultured under flow conditions. Padlock probe-based in situ hybridization (g) detected GOT1 (blue dots) and GOT2 transcripts (yellow dots) in BeWo cells. The anchor, confirming positive signals, is shown in pink. Scale bar: 20 µM. In situ hybridization analysis represent GOT1 (h) and GOT2 (i). Data were normalized to ACTB (j). GOT activity (k) increased in cells exposed to shear stress compared to static conditions. SLC7A8 mRNA (l) increase in cells exposed to shear stress. Data are presented as mean ± SEM based on experiments using five different cell passages. In situ hybridization was performed three times. Outlier were detected with Grubbs’ α = 0.05, normal distribution of data was analyzed with Shapiro–Wilk test. Statistical analysis was performed using one sample t-test. Statistical significance was set at p < 0.05