Fig. 3

TGF-HH axis suppressed ERK1/2 signaling in RS218 infected hBMECs via modulating miR155 and KRAS. A Western blot and qPCR detecting expression of KRAS in RS218 infected hBMECs. The cells were pretreated with rTGFβ1 (at 50 ng/mL) or GANT61 (at 10 μM). **p < 0.01. The qPCR assays were performed in triplicates, and results are presented as mean ± SEM. B IF assays showing the KRAS expression in BMECs of mice challenged by RS218 with or without rTGFβ1 pre-treatment (at 1 μg/kg). The KRAS was stained in red. CD31 was specifically applied for labeling the micro-vessels in green. The cell nucleus was stained in blue with DAPI. Scale bar = 50 μm. C qPCR detecting MIR155HG and miR-155 expression upon RS218 infection in hBMECs. The cells were pretreated with rTGFβ1 (at 50 ng/mL) or GANT61 (at 10 μM). **p < 0.01. The qPCR assays were performed in triplicates, and results are presented as mean ± SEM. D Western blot detecting expression of E-selectin, KRAS and ERK1/2 phosphorylation in RS218 infected hBMECs with or without rTGFβ1 pre-treatment (at 50 ng/mL). The cells were transfected with has-miR-155 mimics, mimics NC, has-miR-155 inhibitors, or inhibitors NC at 50 nM as indicated. E qPCR detecting expression alterations of IL-6, MIP-2, and E-selectin in RS218 infected hBMECs with or without rTGFβ1 pre-treatment (at 50 ng/mL). The cells were transfected with has-miR-155 mimics, mimics NC, has-miR-155 inhibitors, or inhibitors NC at 50 nM as indicated. **p < 0.01. The qPCR assays were performed in triplicates, and results are presented as mean ± SEM