Fig. 7

FSS affects GLUT1, GOT1 and GOT2 expression in different trophoblast layers. First trimester villi were cultured either under static or flow conditions for 24 h. Western Blot for GLUT1 receptor in first trimester placental villi (a) and related band densitometry (b). Key glycolytic enzymes HK2 (c) and PFKP (d) expression levels Representative Western Blot (e) and band densitometry (f) showed no change in placental GOT1 levels. Representative immunofluorescence staining of human placental first trimester explant cultures stained for CK7 (red, upper panel), GOT1 (yellow) (g). Nuclei were stained with DAPI. Software-based image analysis showed downregulation of GOT1 in the CT (h), as well as in the SCT layer (i), and total villous trophoblast compartment (j) under fluidic flow. Immunofluorescence staining of first trimester placental villi for CK7 (red), and GOT2 (yellow) (k). Software-based image analysis showed no significant change of GOT2 intensity in the CT (l), whereas GOT2 intensity increased in the SCT (m) and the total villous trophoblast (n) under fluidic flow. Representative Western Blot (o) and band densitometry (p) showed GOT2 upregulation under flow conditions in placental explants. Immunofluorescence and Western Blot analysis were performed with four different placental samples. 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 for the protein analysis. Statistical analysis for the immunofluorescent staining was performed with an unpaired t-test. Statistical significance was set at p < 0.05. Values represent mean ± SEM.