Fig 2: The significance of cytosolic Notch1 in quiescent HSCs and Rock in activated HSCs. (A) While Notch1 and its target gene, Hes1, are downregulated during HSC activation, the expression of Notch2, a Yap target gene, is strongly upregulated. (B) In contrast to Yap, the Notch1 protein is predominantly present in HSCs (d1) as the Notch1 intracellular domain (N1ICD; Cell Signaling SC-6014). Full-length Notch1 was detected at 300 kDa. The Yap and γ-tubulin loading control blots have been trimmed to remove d1 and d4 (dashed line) (n = 2). (C) Full-length Notch1 and N1ICD are strikingly localized in the cytosol of HSCs (d2) using a cell signaling antibody (#4380). (D) Rock inhibition with 10 µM of Y-27632 (#S1049, Selleckchem) blocked culture-induced HSC activation, most likely by inhibiting the Rock-mediated morphological transition of HSCs. Untreated cells exhibit the cell shape of activated HSCs, which are much larger and contain more α-Sma, whereas Y-27632-treated cells are still stellate cells at d4 with Gfap-containing processes that more closely resemble quiescent HSCs. “d” stands for “day”.

Fig 3: The significance of cytosolic Notch1 in quiescent HSCs and Rock in activated HSCs. (A) While Notch1 and its target gene, Hes1, are downregulated during HSC activation, the expression of Notch2, a Yap target gene, is strongly upregulated. (B) In contrast to Yap, the Notch1 protein is predominantly present in HSCs (d1) as the Notch1 intracellular domain (N1ICD; Cell Signaling SC-6014). Full-length Notch1 was detected at 300 kDa. The Yap and γ-tubulin loading control blots have been trimmed to remove d1 and d4 (dashed line) (n = 2). (C) Full-length Notch1 and N1ICD are strikingly localized in the cytosol of HSCs (d2) using a cell signaling antibody (#4380). (D) Rock inhibition with 10 µM of Y-27632 (#S1049, Selleckchem) blocked culture-induced HSC activation, most likely by inhibiting the Rock-mediated morphological transition of HSCs. Untreated cells exhibit the cell shape of activated HSCs, which are much larger and contain more α-Sma, whereas Y-27632-treated cells are still stellate cells at d4 with Gfap-containing processes that more closely resemble quiescent HSCs. “d” stands for “day”.

Fig 4: The significance of cytosolic Notch1 in quiescent HSCs and Rock in activated HSCs. (A) While Notch1 and its target gene, Hes1, are downregulated during HSC activation, the expression of Notch2, a Yap target gene, is strongly upregulated. (B) In contrast to Yap, the Notch1 protein is predominantly present in HSCs (d1) as the Notch1 intracellular domain (N1ICD; Cell Signaling SC-6014). Full-length Notch1 was detected at 300 kDa. The Yap and γ-tubulin loading control blots have been trimmed to remove d1 and d4 (dashed line) (n = 2). (C) Full-length Notch1 and N1ICD are strikingly localized in the cytosol of HSCs (d2) using a cell signaling antibody (#4380). (D) Rock inhibition with 10 µM of Y-27632 (#S1049, Selleckchem) blocked culture-induced HSC activation, most likely by inhibiting the Rock-mediated morphological transition of HSCs. Untreated cells exhibit the cell shape of activated HSCs, which are much larger and contain more α-Sma, whereas Y-27632-treated cells are still stellate cells at d4 with Gfap-containing processes that more closely resemble quiescent HSCs. “d” stands for “day”.

Fig 5: The significance of cytosolic Notch1 in quiescent HSCs and Rock in activated HSCs. (A) While Notch1 and its target gene, Hes1, are downregulated during HSC activation, the expression of Notch2, a Yap target gene, is strongly upregulated. (B) In contrast to Yap, the Notch1 protein is predominantly present in HSCs (d1) as the Notch1 intracellular domain (N1ICD; Cell Signaling SC-6014). Full-length Notch1 was detected at 300 kDa. The Yap and γ-tubulin loading control blots have been trimmed to remove d1 and d4 (dashed line) (n = 2). (C) Full-length Notch1 and N1ICD are strikingly localized in the cytosol of HSCs (d2) using a cell signaling antibody (#4380). (D) Rock inhibition with 10 µM of Y-27632 (#S1049, Selleckchem) blocked culture-induced HSC activation, most likely by inhibiting the Rock-mediated morphological transition of HSCs. Untreated cells exhibit the cell shape of activated HSCs, which are much larger and contain more α-Sma, whereas Y-27632-treated cells are still stellate cells at d4 with Gfap-containing processes that more closely resemble quiescent HSCs. “d” stands for “day”.