Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Site-1 Protease-Derived Soluble (Pro)Renin Receptor Contributes to Angiotensin II–Induced Hypertension in Mice

doi: 10.1161/HYPERTENSIONAHA.120.15100

Figure Lengend Snippet: Effect of S1P (site-1 protease) inhibition on Ang II (angiotensin II)–induced hypertension in mice. B6129SF1/J mice were randomly divided into the following 4 groups: (1) Control (CTR), (2) Ang II, (3) Ang II + PF, or (4) Ang II + PF + sPRR (soluble [pro]renin receptor)-His. Radiotelemetry was performed to monitor mean arterial blood pressure (MAP; A ), systolic blood pressure (SBP; B ), diastolic blood pressure (DBP; C ), and heart rate (HR; D ) over 6 d. & P <0.05, the interaction detected by repeated-measures ANOVA. * P <0.05 vs CTR for the corresponding period (Bonferroni test). # P <0.05 vs Ang II for the corresponding period (Bonferroni test). $ P <0.05 vs Ang II + PF for the corresponding period (Bonferroni test). Data are mean ± SE. N=4–8 mice per group.

Article Snippet: B6129SF1/J mice were infused for 6 days with control, Ang II (Ang II at 300 ng/kg per day), Ang II + 4-[(Diethylamino)methyl]-N-[2-(2-methoxyphenyl)ethyl]-N-(3R)-3-pyrrolidinyl-benzamide dihydrochloride (PF) (PF at 20 mg/kg per day; HY-13447A, Medchem Express) via a subcutaneously implanted minipump (Alzet model 1007D, Alza) or in conjunction with intravenous sPRR-His infusion at 30 μg/kg per day via jugular vein catheterization connected to a separate osmotic minipump as previously reported.

Techniques: Inhibition, Control

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Site-1 Protease-Derived Soluble (Pro)Renin Receptor Contributes to Angiotensin II–Induced Hypertension in Mice

doi: 10.1161/HYPERTENSIONAHA.120.15100

Figure Lengend Snippet: Assessment of urinary renin levels in Ang II (angiotensin II) infused mice during S1P (site-1 protease) inhibition alone or supplemented with sPRR (soluble [pro]renin receptor)-His. B6129SF1/J mice were randomly divided into the following 4 groups: (1) Control (CTR), (2) Ang II, (3) Ang II + PF, and (4) Ang II + PF + sPRR-His. Urine samples were assayed for renin activity ( A ), prorenin content ( B ), active renin content ( C ), and total renin concentration ( D ). The values were normalized by creatinine. Data are mean ± SE. N=5–10 mice per group. * P <0.05 vs CTR. # P <0.05 vs Ang II. $ P <0.05 vs Ang II + PF.

Article Snippet: B6129SF1/J mice were infused for 6 days with control, Ang II (Ang II at 300 ng/kg per day), Ang II + 4-[(Diethylamino)methyl]-N-[2-(2-methoxyphenyl)ethyl]-N-(3R)-3-pyrrolidinyl-benzamide dihydrochloride (PF) (PF at 20 mg/kg per day; HY-13447A, Medchem Express) via a subcutaneously implanted minipump (Alzet model 1007D, Alza) or in conjunction with intravenous sPRR-His infusion at 30 μg/kg per day via jugular vein catheterization connected to a separate osmotic minipump as previously reported.

Techniques: Inhibition, Control, Activity Assay, Concentration Assay

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Site-1 Protease-Derived Soluble (Pro)Renin Receptor Contributes to Angiotensin II–Induced Hypertension in Mice

doi: 10.1161/HYPERTENSIONAHA.120.15100

Figure Lengend Snippet: Assessment of renal inner medullary renin levels and AGT levels in Ang II (angiotensin II) infused mice during S1P (site-1 protease) inhibition alone or supplemented with sPRR (soluble [pro]renin receptor)-His. B6129SF1/J mice were randomly divided into the following 4 groups: (1) Control (CTR), (2) Ang II, (3) Ang II + PF, and (4) Ang II + PF + sPRR-His. The lysates of the renal inner medulla were assayed for renin activity ( A ), prorenin content ( B ), active renin content ( C ), and total renin concentration ( D ). The values were normalized by protein content. Data are mean ± SE. N=5–10 mice per group. * P <0.05 vs CTR. # P <0.05 vs Ang II. $ P <0.05 vs Ang II + PF. The renal inner medulla was also subjected to quantitative real-time polymerase chain reaction (qRT-PCR) analysis of renin mRNA expression normalized by GAPDH ( E ). Data are mean ± SE. N=5–10 per group. * P <0.05 vs CTR. # P <0.05 vs Ang II. $ P <0.05 vs Ang II + PF.

Article Snippet: B6129SF1/J mice were infused for 6 days with control, Ang II (Ang II at 300 ng/kg per day), Ang II + 4-[(Diethylamino)methyl]-N-[2-(2-methoxyphenyl)ethyl]-N-(3R)-3-pyrrolidinyl-benzamide dihydrochloride (PF) (PF at 20 mg/kg per day; HY-13447A, Medchem Express) via a subcutaneously implanted minipump (Alzet model 1007D, Alza) or in conjunction with intravenous sPRR-His infusion at 30 μg/kg per day via jugular vein catheterization connected to a separate osmotic minipump as previously reported.

Techniques: Inhibition, Control, Activity Assay, Concentration Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Site-1 Protease-Derived Soluble (Pro)Renin Receptor Contributes to Angiotensin II–Induced Hypertension in Mice

doi: 10.1161/HYPERTENSIONAHA.120.15100

Figure Lengend Snippet: Assessment of renal regional expression of ENaC subunits in Ang II (angiotensin II) infused mice during S1P (site-1 protease) inhibition alone or supplemented with sPRR (soluble [pro]renin receptor)-His. B6129SF1/J mice were randomly divided into the following 4 groups: (1) Control (CTR), (2) Ang II, (3) Ang II + PF, and (4) Ang II + PF + sPRR-His. The renal cortex and the renal medulla were subjected to quantitative real-time polymerase chain reaction (qRT-PCR) analysis of mRNA expression of α-ENaC ( Α and D ), β-ENaC ( Β and E ), and γ-ENaC ( C and F ). A–C , renal cortex. D–F , renal medulla. The mRNA expression was normalized by GAPDH. Data are mean ± SE. N=5–10 mice per group. * P <0.05 vs CTR. # P <0.05 vs Ang II. $ P <0.05 vs Ang II + PF.

Article Snippet: B6129SF1/J mice were infused for 6 days with control, Ang II (Ang II at 300 ng/kg per day), Ang II + 4-[(Diethylamino)methyl]-N-[2-(2-methoxyphenyl)ethyl]-N-(3R)-3-pyrrolidinyl-benzamide dihydrochloride (PF) (PF at 20 mg/kg per day; HY-13447A, Medchem Express) via a subcutaneously implanted minipump (Alzet model 1007D, Alza) or in conjunction with intravenous sPRR-His infusion at 30 μg/kg per day via jugular vein catheterization connected to a separate osmotic minipump as previously reported.

Techniques: Expressing, Inhibition, Control, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Site-1 Protease-Derived Soluble (Pro)Renin Receptor Contributes to Angiotensin II–Induced Hypertension in Mice

doi: 10.1161/HYPERTENSIONAHA.120.15100

Figure Lengend Snippet: Assessment of renal protein abundances of α-ENaC and PRR ([pro]renin receptor)/sPRR (soluble PRR), and urinary sPRR excretion in Ang II (angiotensin II) infused mice during S1P (site-1 protease) inhibition alone or supplemented with sPRR-His. The renal cortex ( A and B ) and inner medulla ( C and D ) from mice treated with control (CTR), Ang II, Ang II + PF, or Ang II + PF + sPRR-His were subjected to immunoblotting analysis of protein expression α-ENaC and PRR/sPRR. The protein abundances were analyzed by densitometry and normalized by β-actin. The values were shown underneath the blots. Data are mean ± SE. N=5–10 per group. * P <0.05 vs CTR. # P <0.05 vs Ang II. $ P <0.05 vs Ang II + PF. Urine samples were subjected to ELISA measurement of urinary sPRR concertation. The values were expressed as 24 h urine output of sPRR. Data are mean ± SE. N=6 mice per group. * P <0.05 vs CTR. # P <0.05 vs Ang II.

Article Snippet: B6129SF1/J mice were infused for 6 days with control, Ang II (Ang II at 300 ng/kg per day), Ang II + 4-[(Diethylamino)methyl]-N-[2-(2-methoxyphenyl)ethyl]-N-(3R)-3-pyrrolidinyl-benzamide dihydrochloride (PF) (PF at 20 mg/kg per day; HY-13447A, Medchem Express) via a subcutaneously implanted minipump (Alzet model 1007D, Alza) or in conjunction with intravenous sPRR-His infusion at 30 μg/kg per day via jugular vein catheterization connected to a separate osmotic minipump as previously reported.

Techniques: Inhibition, Control, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: Site-1 Protease-Derived Soluble (Pro)Renin Receptor Contributes to Angiotensin II–Induced Hypertension in Mice

doi: 10.1161/HYPERTENSIONAHA.120.15100

Figure Lengend Snippet: Role of S1P (site-1 protease)-derived sPRR (soluble [pro]renin receptor) in mediation of Ang II (angiotensin II)–induced ENaC activation in vitro. Confluent mouse cortical collecting duct cell line (mpkCCD) cells grown on Snapwells were pretreated with 10 μmol/L PF for 1 h and then treated with 1 μmol/L Ang II and 10 nmol/L sPRR-His for 24 h. Amiloride-sensitive transepithelial Na + transport, an index of ENaC activity, was recorded by the Ussing chamber technique. Medium sPRR was measured by using ELISA. A , ENaC activity in CTR, Ang II, Ang II + PF, and Ang II + PF + sPRR-His groups. Data are mean ± SE. N=4 per group. * P <0.05 vs CTR. # P <0.05 vs Ang II. $ P <0.05 vs Ang II + PF. B , Medium sPRR content in cells treated with CTR, Ang II, or Ang II + PF were determined by using ELISA. Data are mean ± SE. N=4 per group. * P <0.05 vs CTR. # P <0.05 vs Ang II. C , The cells were transfected with S1P siRNA or scrambled siRNA, followed by 24-h Ang II treatment and Ussing chamber measurement of ENaC activity. Date are mean ± SE. N=4 per group. * P <0.05 vs scrambled siRNA. D , The cells were pretreated with 1 μmol/L Aliskiren for 0.5 h or 10 μmol/L PF for 1 h, followed by 24-h Ang II treatment and Ussing chamber measurement of ENaC activity. Date are mean ± SE. N=4 per group. * P <0.05 vs CTR. # P <0.05 vs Ang II.

Article Snippet: B6129SF1/J mice were infused for 6 days with control, Ang II (Ang II at 300 ng/kg per day), Ang II + 4-[(Diethylamino)methyl]-N-[2-(2-methoxyphenyl)ethyl]-N-(3R)-3-pyrrolidinyl-benzamide dihydrochloride (PF) (PF at 20 mg/kg per day; HY-13447A, Medchem Express) via a subcutaneously implanted minipump (Alzet model 1007D, Alza) or in conjunction with intravenous sPRR-His infusion at 30 μg/kg per day via jugular vein catheterization connected to a separate osmotic minipump as previously reported.

Techniques: Derivative Assay, Activation Assay, In Vitro, Activity Assay, Enzyme-linked Immunosorbent Assay, Transfection

Journal: Cell reports

Article Title: SREBP-1 upregulates SOAT1 to promote tumor growth by preventing lipotoxicity

doi: 10.1016/j.celrep.2025.116896

Figure Lengend Snippet: (A) Schematic of the process of SREBP-1 activation and its potential role in regulating SOAT1 expression. The inhibitor fatostatin blocks trafficking of the SCAP/SREBP-1 complex, and PF429242 suppresses SREBP-1 cleavage by inhibiting site 1 protease (S1P). The diagram was created using BioRender. (B–G) Western blot analysis of the indicated proteins (B, C, and F) and representative fluorescence images of LDs stained with BODIPY 493/503 (D, E, and G) in GBM or lung cancer cells following treatment with fatostain (Fato) (B and D) or PF429242 (C and E) for 24 h or SREBP-1 knockdown using lentivirus-mediated shRNA for 48 h (F and G). (H) Schematic of the putative SREBP-1 binding sites (SREs) and a negative control (NC) region in the SOAT1 promoter (top). Arrows indicate the positions of primers used for PCR analysis following chromatin immunoprecipitation (ChIP) with either control immunoglobulin G (IgG) or anti-SREBP-1 antibody (bottom). (I) Measuring the activities of the SOAT1 promoters using a pGL3-luciferase (Luc) reporter plasmid in HEK293-FT cells in response to the expression of N -terminal SREBP-1a or -1c isoforms for 48 h (J) Mutagenesis of the putative SRE in SOAT1 promoter and measuring its activity in response to expression of N-terminal SREBP-1a or -1c isoforms for 48 h (K) Real-time PCR analysis of SOAT1 gene expression in GBM30 or H1299 cells following adenoviral expression of FLAG-tagged N-terminal SREBP-1a (Ad- 1aN ), or SREBP-1c (Ad- 1cN ) isoforms compared to a control vector (Ad- null ) for 48 h (L and M) Western blot (L) and fluorescence imaging of LDs stained with BODIPY 493/503 (M) in GBM30 and H1299 cells after adenoviral expression of FLAG-tagged N-terminal SREBP-1a or -1c isoforms for 48 h under 5% FBS conditions. All experiments were performed in three independent biological replicates. For (D), (E), (G)–(K), and (M), data are presented as means ± SD. Nuclei were counterstained with Hoechst 33342 (blue). Scale bar, 10 μm. Quantification of LDs was performed using ImageJ from >30 cells, for (D), (E), (G), and (M). Statistical significance was analyzed by one-way ANOVA compared with vehicle-treated controls for (G) or Student’s t test for (D), (E), (H), (I)–(K), and (M). See also .

Article Snippet: PF429242 , Selleckchem.com , Cat# S6418.

Techniques: Activation Assay, Expressing, Western Blot, Fluorescence, Staining, Knockdown, shRNA, Binding Assay, Negative Control, Chromatin Immunoprecipitation, Control, Luciferase, Plasmid Preparation, Mutagenesis, Activity Assay, Real-time Polymerase Chain Reaction, Gene Expression, Imaging

Journal: Journal of Translational Medicine

Article Title: Cathepsin H as a causal risk factor and therapeutic target in proliferative diabetic retinopathy

doi: 10.1186/s12967-025-07314-4

Figure Lengend Snippet: Docking results and interactions of cathepsin H with ( A ) SSR 69071 ( B ) PF429242 , in 3D and 2D formats

Article Snippet: Molecular dynamics simulations (100 ns) of the SSR 69071-CtsH and PF429242 -CtsH complexes confirmed stable dynamics: root mean square deviation (RMSD) plateaued below 0.2 nm after 40 ns (Fig. A), indicating conformational stability.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Cathepsin H as a causal risk factor and therapeutic target in proliferative diabetic retinopathy

doi: 10.1186/s12967-025-07314-4

Figure Lengend Snippet: Effects of PF429242 and SSR 69071 on HUVEC healing rate, and lysosomal function. ( A ) Representative images of HUVECs at 0, 12, and 24 hours after treatment with different concentrations of PF429242 (veh, 2.5, 5, 10 μM); ( B ) Healing rate of HUVECs at 0 and 12 hours after treatment with different concentrations of PF429242 (veh, 2.5, 5, 10 μM); ( C ) Healing rate of HUVECs at 0 and 24 hours after treatment with different concentrations of PF429242 (veh, 2.5, 5, 10 μM); ( D ) Representative images of HUVECs at 0, 12, and 24 hours after treatment with different concentrations of SSR 69071 (veh, 0.01, 0.1, 1 μM); ( E ) Healing rate of HUVECs at 0 and 12 hours after treatment with different concentrations of SSR 69071 (0, 0.01, 0.1, 1 μM); ( F ) Healing rate of HUVECs at 0 and 24 hours after treatment with different concentrations of SSR 69071 (0, 0.01, 0.1, 1 μM); ( G ) Representative images of HUVECs treated with 10 μM PF429242 or 1 μM SSR 69071 for 4 hours, detected by LysoTracker Red staining (nuclei counterstained with Hoechst 33,342). ( H ) Representative images of HUVECs treated with 10 μM PF429242 or 1 μM SSR 69071 for 4 hours, captured by a Multi-SIM system, with lysosomes stained by LysoTracker Red and nuclei counterstained with Hoechst 33342. ( I ) Lysosomal area per cell of HUVECs treated with 10 μM PF429242 or 1 μM SSR 69071 for 4 hours. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Molecular dynamics simulations (100 ns) of the SSR 69071-CtsH and PF429242 -CtsH complexes confirmed stable dynamics: root mean square deviation (RMSD) plateaued below 0.2 nm after 40 ns (Fig. A), indicating conformational stability.

Techniques: Staining