TGF-beta mediated hepatic stellate cells activation is dampened in the absence of EPHB2 in vitro.

Hepatic stellate cells (HSCs) are the major effector cell-type responsible for the production of collagens in hepatic fibrogenesis. In this study we showed that when EPHB2 is silenced in human primary HSCs, they become refractory to TGFbeta-induced-HSCs-to-myofibroblasts transdifferentiation. More importantly, EphB2-specific deletion in HSCs using the tamoxifen inducible PdgfCre-eRT2 promoer driver is sufficient to mitigate MASH fibrosis in mice. Overall design: Human hepatic stellate cells (HSCs) (70% confluency) were transfected with EPHB2 Mission®-esiRNA (Sigma Aldrich, USA) delivered into the cells using the lipofectamine® RNAiMAX (Invitrogen, USA) according to the manufacturer's instruction. Mission®-esiRNA complexes targeting eGFP were used as nontargeting control. After 48H incubation, transfected HSCs were stimulated with recombinant human TGF-ß1 at 10 ng/ml for another 24 hour. Cells were harvested, total RNA was extracted using RNA Stat60® (Tel-Test Inc, USA) following standard phenol-chloroform procedure.