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Impact of Fasudil and Y-27632 on HIF-1α and Rho-Kinase Activity Under Hypoxic Conditions

This study explores the effects of Fasudil and Y-27632 on HIF-1α levels and Rho-kinase activity under varying hypoxic conditions. Supplemental Figures provide detailed assays showing the modulation of HIF-1α expression and its polyubiquitination, along with Rho-kinase signaling. Assessments were made using inhibitors and other reagents to establish the connections between these regulatory pathways. The findings shed light on the therapeutic potential of targeting Rho-kinase in diseases associated with hypoxia.

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Impact of Fasudil and Y-27632 on HIF-1α and Rho-Kinase Activity Under Hypoxic Conditions

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  1. Supplemental Figure 1. a db/mdb/dbdb/db + fasudil b db/mdb/dbdb/db + fasudil c db/m db/db db/db + fasudil NS

  2. Supplemental Figure 2. Hypoxia － ＋ － ＋ Y-27632 － － ＋ ＋

  3. Supplemental Figure 3. a b c DFX DFX DFX DFX DFX DFX DFX － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ Y-27632 Y-27632 Y-27632 Y-27632 Y-27632 Y-27632 Y-27632 － － ＋ ＋ － － ＋ ＋ － － ＋ ＋ － － ＋ ＋ － － ＋ ＋ － － ＋ ＋ － － ＋ ＋ d e f g h ** IB: HIF-1α IB: β-actin DFX － ＋ － ＋ － ＋ Y-27632 － － ＋ ＋ － ＋ MG132 － － － － ＋ ＋ ** ** ** ** ** ** ** ** ** ** ** ** ** **

  4. Supplemental Figure 4. Polyubiquitinated HIF-1α IP:HIF-1α IB:Ub IB:HIF-1α IB: β-actin Y-27632 － ＋ MG132 ＋ ＋

  5. Supplemental Figure 5. Rho-kinase activity IB: p-MYPT1 IB: MYPT1 DFX － ＋

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