Journal: International journal of molecular sciences

Article Title: Hepatocyte Specific gp130 Signalling Underlies APAP Induced Liver Injury.

doi: 10.3390/ijms23137089

Figure Lengend Snippet: Figure 2. Hepatic CYP2E1 and glutathione levels and serum and molecular markers of liver injury in mice with hepatocyte-specific gp130 deletion and control mice. (A) Representative immunohisto- chemistry images (scale bars, 200 µm; representative dataset from n = 3 mice/group). (B) Schematic of APAP induction in wild-type (WT) and CKOgp130 mice: overnight-fasted mice were injected with saline or APAP (400 mg kg−1); livers and serum were collected either 0.5, 6, or 24 h post-APAP dosing for experiments shown in (C–F) and Figure 3. (C) Hepatic GSH concentrations at 0.5 and 6 h post-APAP. (D) ALT, (E) AST, and (F) Western blots of IL11, NOX4, GAPDH, p-ERK, ERK, p-JNK, JNK, p-STAT3, STAT3, Cl. CASP3, and CASP3 (n = 3/group) at 6 h post-APAP. (G) Representa- tive H&E-stained liver images (scale bars, 200 µm) at 6 h post-APAP dosing and quantification of liver necrotic area. (C–E,G) Data are shown in a box-and-whisker plot with median (middle line), 25th–75th percentiles (box) and min-max values (whiskers), with a 2-way ANOVA with Sidak’s correction; 0.5 h time point: saline (n = 4 mice/genotype), APAP (n = 4 mice/genotype), 6: saline (n = 5 mice/genotype), APAP (n = 6 mice/genotype).

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) and Colorimetric Assays The levels of IL11 in mouse serum were quantified using Mouse IL11 DuoSet ELISA (DY418, R&D Systems, Minneapolis, MN, USA).

Techniques: Control, Immunohistochemistry, Injection, Saline, Western Blot, Staining, Whisker Assay

Journal: International journal of molecular sciences

Article Title: Hepatocyte Specific gp130 Signalling Underlies APAP Induced Liver Injury.

doi: 10.3390/ijms23137089

Figure Lengend Snippet: Figure 3. Hepatocyte-specific deletion of gp130 limits APAP-induced liver injury and promotes hepatic regeneration 24 h after injury. (A) Serum IL11 levels, (B) serum ALT levels, (C) serum AST levels, (D) hepatic GSH levels, (E) Western blots showing hepatic levels of IL11, PCNA, Cyclin D1, and GAPDH as internal contro (n = 3/group)l, (F) hepatic mRNA expression of pro-inflammatory markers (Ccl2, Ccl5, Il1β, Il6, Tnfα), (G) Western blots showing hepatic levels of p-ERK, ERK, p-JNK, JNK, Cl. CASP3, CASP3, p-STAT3, and STAT3 (n = 3/group), (H) representative H&E-stained liver images (scale bars, 200 µm) and quantification of liver necrotic area, (I) IHC staining of Ki67 in the livers of mice (scale bars, 200 µm) and quantification of Ki67+ve cells for experiments shown in Figure 2B. (A–D,F,H,I) Saline (n = 5 mice/genotype), APAP (n = 6 mice/genotype); data are shown in a box-and-whisker plot with median (middle line), 25th–75th percentiles (box), and min-max values (whiskers), with a 2-way ANOVA with Sidak’s correction.

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) and Colorimetric Assays The levels of IL11 in mouse serum were quantified using Mouse IL11 DuoSet ELISA (DY418, R&D Systems, Minneapolis, MN, USA).

Techniques: Western Blot, Expressing, Staining, Immunohistochemistry, Saline, Whisker Assay

Journal: International journal of molecular sciences

Article Title: Hepatocyte Specific gp130 Signalling Underlies APAP Induced Liver Injury.

doi: 10.3390/ijms23137089

Figure Lengend Snippet: Figure 4. Generation and validation of hepatocyte-specific Il11-deficient adult mice. (A) Schematic design showing the deletion of exons 2–5 of the mouse Il11 locus and the gene targeting strategy. (B) Representative genotyping of Il11+/+, Il11loxP/+, or Il11loxP/loxP mice. (C) Schematic showing AAV8-induced hepatocyte-specific Il11 knockout mice (CKOIl11). (D) Il11 mRNA (n = 5/group) and (E) IL11 protein (n = 3/group) expression in the heart, liver, lung, and kidney from WT and CKOIl11

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) and Colorimetric Assays The levels of IL11 in mouse serum were quantified using Mouse IL11 DuoSet ELISA (DY418, R&D Systems, Minneapolis, MN, USA).

Techniques: Biomarker Discovery, Knock-Out, Expressing

Journal: International journal of molecular sciences

Article Title: Hepatocyte Specific gp130 Signalling Underlies APAP Induced Liver Injury.

doi: 10.3390/ijms23137089

Figure Lengend Snippet: Figure 5. Hepatic CYP2E1 and glutathione levels and serum and molecular markers of liver injury in mice with hepatocyte-specific Il11 deletion and control mice. (A) Representative immunohisto- chemistry images (scale bars, 200 µm; representative dataset from n = 3 mice/group). (B) Schematic of induction of APAP injury in wild-type (WT) and CKOIl11 mice: overnight-fasted mice were intraperitoneally injected with saline or APAP (400 mg kg−1); livers and serum were collected either 0.5, 6, or 24 h following APAP administration for experiments shown in (C–F) and Figure 6. (C) Hepatic GSH concentrations at 0.5 and 6 h post-APAP. (D) ALT, (E) AST, and (F) Western blots of IL11, NOX4, GAPDH, p-ERK, ERK, p-JNK, JNK, Cl. CASP3, and CASP3 at 6 h post-APAP. (G) Representative H&E-stained liver images (scale bars, 200 µm) at 6 h post-APAP dosing quan- tification of the liver necrotic area. (C–E,G) Data are shown in a box-and-whisker plot with median (middle line), 25th–75th percentiles (box), and min-max values (whiskers), with a 2-way ANOVA with Sidak’s correction; 0.5 h time point: saline (n = 4 mice/genotype), APAP (n = 4 mice/genotype), 6 h time point: saline (n = 5 mice/genotype), APAP (n = 6 mice/genotype).

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) and Colorimetric Assays The levels of IL11 in mouse serum were quantified using Mouse IL11 DuoSet ELISA (DY418, R&D Systems, Minneapolis, MN, USA).

Techniques: Control, Immunohistochemistry, Injection, Saline, Western Blot, Staining, Whisker Assay

Journal: International journal of molecular sciences

Article Title: Hepatocyte Specific gp130 Signalling Underlies APAP Induced Liver Injury.

doi: 10.3390/ijms23137089

Figure Lengend Snippet: Figure 6. Hepatocyte-specific deletion of Il11 reduces APAP-induced liver injury and promotes hepatic regeneration 24 h after injury. (A) Serum IL11 levels, (B) Western blots showing hepatic levels of IL11, PCNA, Cyclin D1, and GAPDH as the internal control (n = 3/group), (C) serum ALT levels, (D) serum AST levels, (E) hepatic GSH levels, (F) hepatic mRNA expression of pro-inflammatory markers (Ccl2, Ccl5, Il1β, Il6, Tnfα), (G) Western blots showing hepatic levels of p-ERK, ERK, p-JNK, JNK, Cl. CASP3, CASP3, p-STAT3, and STAT3 (n = 3/group), (H) representative H&E-stained liver images (scale bars, 200 µm) and quantification of the liver necrotic area, (I) IHC staining of Ki67 in the livers of mice (scale bars, 200 µm) and quantification of Ki67+ve cells for experiments shown in Figure 5B. (A,C–F,H,I) Saline (n = 5 mice/genotype), APAP (n = 6 mice/genotype); data are shown in a box-and-whisker plot with median (middle line), 25th–75th percentiles (box), and min-max values (whiskers), with a 2-way ANOVA with Sidak’s correction.

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) and Colorimetric Assays The levels of IL11 in mouse serum were quantified using Mouse IL11 DuoSet ELISA (DY418, R&D Systems, Minneapolis, MN, USA).

Techniques: Western Blot, Control, Expressing, Staining, Immunohistochemistry, Saline, Whisker Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Krüppel‐Like Factor 15/Interleukin 11 Axis‐Mediated Adventitial Remodeling Depends on Extracellular Signal‐Regulated Kinases 1 and 2 Activation in Angiotensin II–Induced Hypertension

doi: 10.1161/JAHA.120.020554

Figure Lengend Snippet: A , Representative immunofluorescence staining and quantitative analysis of IL‐11 (red), α‐smooth muscle actin (ACTA2) (green) in thoracic aorta. The 4′,6‐diamidino‐2‐phenylindole was used to stain the nucleus in blue. Bar=50 µm. B , IL‐11 mRNA was stained with fluorescence by situ hybridization assay. Bar=50 µm. C , The mRNA level of IL‐11 (1.00‐ and 12.47‐fold) in thoracic aorta of wild‐type mice with or without Ang II infusion was quantified by quantitative reverse transcription–polymerase chain reaction (RT‐qPCR). D , The media (M) and adventitia (A) were treated with Ang II (10 −6 mol/L) for 4 hours. The mRNA level of IL‐11 was quantified by RT‐qPCR (1.00‐, 8.23‐, 2.18‐, and 15.4‐fold). I indicates intima. M indicates media; and A indicates adventitia. ** P <0.01, *** P < 0.001.

Article Snippet: For Ang II+IL‐11 neutralizing antibody group, wild‐type (WT) mice were treated with 1 mg/kg intravenous IL‐11 neutralizing antibody (R&D; MAB418) at 7, 9, 11, and 13 days after Ang II infusion.

Techniques: Immunofluorescence, Staining, Fluorescence, Hybridization, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Krüppel‐Like Factor 15/Interleukin 11 Axis‐Mediated Adventitial Remodeling Depends on Extracellular Signal‐Regulated Kinases 1 and 2 Activation in Angiotensin II–Induced Hypertension

doi: 10.1161/JAHA.120.020554

Figure Lengend Snippet: A , Representative hematoxylin and eosin (H&E) and picrosirius red staining of the thoracic aorta tissues in wild type (WT) and IL‐11‐/‐ mice after Ang II infusion for 14 days. Bar=50 µm. B through D , Quantitative analysis of wall thickness ( B ), wall area ( C ), and collagen area ( D ). E , Systolic blood pressure of the 4 groups. F , Representative immunofluorescence staining and quantitative analysis of ER‐TR7 (red), α‐smooth muscle actin (ACTA2) (green), CD68 (green), and Krüppel‐like factor 15 (KLF15) (red) in thoracic aorta. The 4′,6‐diamidino‐2‐phenylindole was used to stain the nucleus in blue. Bar=50 µm. G , The mRNA levels of KLF15 (1.00‐, 0.27‐, 1.07‐, and 0.22‐fold), collagen, type I, α 1 (COL1a1) (1.00‐, 14.52‐, 0.43‐, and 1.74‐fold), collagen, type III, α 1 (COL3a1) (1.00‐, 1.69‐, 0.86‐, and 1.17‐fold), interleukin 6 (IL‐6) (1.00‐, 1.93‐, 0.33‐, and 0.43‐fold) and chemokine (C‐C motif) ligand 2 (CCL2) (1.00‐, 2.11‐, 0.42‐, and 0.53‐fold) were measured by quantitative reverse transcription–polymerase chain reaction. A indicates adventitia. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: For Ang II+IL‐11 neutralizing antibody group, wild‐type (WT) mice were treated with 1 mg/kg intravenous IL‐11 neutralizing antibody (R&D; MAB418) at 7, 9, 11, and 13 days after Ang II infusion.

Techniques: Staining, Immunofluorescence, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Krüppel‐Like Factor 15/Interleukin 11 Axis‐Mediated Adventitial Remodeling Depends on Extracellular Signal‐Regulated Kinases 1 and 2 Activation in Angiotensin II–Induced Hypertension

doi: 10.1161/JAHA.120.020554

Figure Lengend Snippet: Wild‐type mice were randomly divided into 4 groups treated with rmIL‐11 or IL‐11 neutralizing antibody with or without Ang II infusion. A , Representative hematoxylin and eosin (H&E) and picrosirius red staining of the thoracic aorta tissues. Bar=50 µm. B through D , Quantitative analysis of wall thickness ( B ), wall area ( C ), and collagen area ( D ). E , Systolic blood pressure of the 4 groups. F , Representative immunofluorescence staining and quantitative analysis of ER‐TR7 (red), α‐smooth muscle actin (ACTA2) (green), CD68 (green), and Krüppel‐like factor 15 (KLF15) (red) in thoracic aorta. The 4′,6‐diamidino‐2‐phenylindole was used to stain the nucleus in blue. Bar=50 µm. G , The mRNA levels of KLF15 (1.00‐, 0.47‐, 0.39‐, and 0.40‐fold), collagen, type I, α 1 (COL1a1) (1.00‐, 2.85‐, 0.81‐, and 3.70‐fold), collagen, type III, α 1 (COL3a1) (1.00‐, 2.69‐, 0.85‐, and 6.67‐fold), interleukin‐6 (IL‐6) (1.00‐, 1.75‐, 0.23‐, and 1.95‐fold), and chemokine (C‐C motif) ligand 2 (CCL2) (1.00‐, 1.83‐, 0.39‐, and 2.76‐fold) were measured by quantitative reverse transcription–polymerase chain reaction. A indicates adventitia. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: For Ang II+IL‐11 neutralizing antibody group, wild‐type (WT) mice were treated with 1 mg/kg intravenous IL‐11 neutralizing antibody (R&D; MAB418) at 7, 9, 11, and 13 days after Ang II infusion.

Techniques: Staining, Immunofluorescence, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Krüppel‐Like Factor 15/Interleukin 11 Axis‐Mediated Adventitial Remodeling Depends on Extracellular Signal‐Regulated Kinases 1 and 2 Activation in Angiotensin II–Induced Hypertension

doi: 10.1161/JAHA.120.020554

Figure Lengend Snippet: A, Adventitial fibroblasts ( AFs) were exposed to angiotensin II (Ang II) (10 −6 mol/L) for indicated time. The protein expressions of IL‐11 (1.00‐, 1.47‐, 2.20‐, 2.58‐, and 2.80‐fold) and IL‐11 receptor subunit α (IL‐11Rα) were quantified by Western blotting. N=3. B, AFs were exposed to Ang II (10 −6 mol/L) for indicated time. The mRNA levels of IL‐11 (1.00‐, 2.73‐, 5.30‐, 3.42‐, 1.99‐, and 1.98‐fold) and IL‐11Rα were measured by quantitative reverse transcription–polymerase chain reaction (RT‐qPCR). N=6. C, AFs were transfected with control adenovirus (AdCON) or adenovirus‐mediated KLF15 (AdKLF15) for 48 hours, then cells were treated with Ang II (10 −6 mol/L) for 4 hours. The mRNA level of IL‐11 (1.00‐, 12.40‐, 0.68‐, and 5.42‐fold) was measured by RT‐qPCR. N=4. D, Human embryonic kidney 293T cells were stimulated with PBS or Ang II (10 −6 mol/L) for 4 hours. The transcription activity of IL‐11 promoter was measured by luciferase assays. N=4. E, AFs were transfected with AdCON or AdKLF15 for 48 hours, then cells were stimulated with Ang II for 24 hours. The protein levels of IL‐11 (1.00‐, 1.65‐, 0.74‐, and 0.90‐fold), collagen, type I, α 1 (COL1a1) (1.00‐, 1.92‐, 0.98‐, and 1.26‐fold), and α‐smooth muscle actin (ACTA2) (1.00‐, 1.88‐, 0.85‐, and 0.80‐fold) were measured by Western blotting. N=3. The right panel was the quantity of the Western blot. F, Cells were transfected with control small interfering RNA (siRNA; siCON) or KLF15 siRNA (siKLF15) for 48 hours, then cells were pretreated with neutralizing antibody 1 hour before Ang II treatment for 24 hours. The protein levels of IL‐11 (1.00‐, 1.46‐, 0.88‐, 1.91‐, and 0.89‐fold), COL1a1 (1.00‐, 2.00‐, 1.00‐, 2.95‐, and 2.07‐fold), and ACTA2 (1.00‐, 1.42‐, 0.97‐, 2.42‐, and 1.02‐fold) were measured by Western blotting. The right panel was the quantity of the Western blot. G, Cells were transfected with siCON or siKLF15 for 48 hours, then cells were pretreated with neutralizing antibody 1 hour before Ang II treatment for 24 hours. The mRNA levels of COL1a1 (1.00‐, 1.62‐, 1.05‐, 2.56‐, and 2.53‐fold) and ACTA2 (1.00‐, 1.44‐, 0.98‐, 2.31‐, and 1.93‐fold) were measured by RT‐qPCR. N=5. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: For Ang II+IL‐11 neutralizing antibody group, wild‐type (WT) mice were treated with 1 mg/kg intravenous IL‐11 neutralizing antibody (R&D; MAB418) at 7, 9, 11, and 13 days after Ang II infusion.

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Activity Assay, Luciferase, Small Interfering RNA

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Krüppel‐Like Factor 15/Interleukin 11 Axis‐Mediated Adventitial Remodeling Depends on Extracellular Signal‐Regulated Kinases 1 and 2 Activation in Angiotensin II–Induced Hypertension

doi: 10.1161/JAHA.120.020554

Figure Lengend Snippet: A , Adventitial fibroblasts ( AFs) were exposed to Ang II (10 −6 mol/L) for indicated time, and the protein level of phosphorylated ERK1/2 (p‐ERK1/2) was measured by Western blotting. B , AFs were pretreated with PBS or IL‐11 neutralizing antibody 1 hour before Ang II (10 −6 mol/L) treatment. Then, AFs were exposed to Ang II or PBS for indicated time. The level of p‐ERK1/2 (1.00‐, 3.14‐, 2.25‐, 0.76‐, 2.74‐, and 0.73‐fold) was measured by Western blotting. N=4. C , AFs were transfected with control small interfering RNA (siRNA; siCON) or Krüppel‐like factor 15 (KLF15) siRNA (siKLF15) for 48 hours. AFs were pretreated by IL‐11 neutralizing antibody 1 hour before they were stimulated by Ang II for 5 minutes (1.00‐, 3.52‐, 4,04‐, 0.88‐, 3.76‐, and 3.57‐fold) or 4 hours (1.00‐, 2.31‐, 1.42‐, 1.08‐, 3.90‐, and 1.46‐fold). p‐ERK1/2 expression was measured by Western blotting. N=4. D , AFs were incubated with ERK1/2 signaling inhibitor (ERKi) 1 hour before or after Ang II 24‐hour stimulation. The protein levels of IL‐11 (1.00‐, 1.41‐, 0.96‐, and 1.26‐fold), KLF15 (1.00‐, 0.62‐, 1.08‐, and 0.49‐fold), collagen, type I, α 1 (COL1a1) (1.00‐, 1.69‐, 0.80‐, and 0.92‐fold), and α‐smooth muscle actin (ACTA2) (1.00‐, 1.53‐, 0.95‐, 0.92‐, and 0.49‐fold) were measured by Western blotting. The right panel was the quantity of the Western blot. N=4. CON indicates control. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: For Ang II+IL‐11 neutralizing antibody group, wild‐type (WT) mice were treated with 1 mg/kg intravenous IL‐11 neutralizing antibody (R&D; MAB418) at 7, 9, 11, and 13 days after Ang II infusion.

Techniques: Western Blot, Transfection, Small Interfering RNA, Expressing, Incubation

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Krüppel‐Like Factor 15/Interleukin 11 Axis‐Mediated Adventitial Remodeling Depends on Extracellular Signal‐Regulated Kinases 1 and 2 Activation in Angiotensin II–Induced Hypertension

doi: 10.1161/JAHA.120.020554

Figure Lengend Snippet: A and B, Representative immunofluorescence staining and quantitative analysis of phosphorylated ERK1/2 (p‐ERK1/2) (green) in thoracic aorta of different groups. The 4′,6‐diamidino‐2‐phenylindole was used to stain the nucleus in blue. Bar=50 µm. A indicates adventitia; IL‐11 –/– , IL‐11 knockout; rmIL‐11, recombination mouse IL‐11; and WT, wild type. ** P < 0.01, *** P < 0.001.

Article Snippet: For Ang II+IL‐11 neutralizing antibody group, wild‐type (WT) mice were treated with 1 mg/kg intravenous IL‐11 neutralizing antibody (R&D; MAB418) at 7, 9, 11, and 13 days after Ang II infusion.

Techniques: Immunofluorescence, Staining, Knock-Out

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Krüppel‐Like Factor 15/Interleukin 11 Axis‐Mediated Adventitial Remodeling Depends on Extracellular Signal‐Regulated Kinases 1 and 2 Activation in Angiotensin II–Induced Hypertension

doi: 10.1161/JAHA.120.020554

Figure Lengend Snippet: A, Adventitial fibroblasts (AFs) were incubated with IL‐11 neutralizing antibody 1 hour before Ang II 24‐hour treatment. The protein levels of collagen, type I, α 1 (COL1a1) (1.00‐, 1.19‐, 2.10‐, and 0.95‐fold), α‐smooth muscle actin (ACTA2) (1.00‐, 1.00‐, 1.49‐, and 0.97‐fold), and Krüppel‐like factor 15 (KLF15) (1.00‐, 0.87‐, 0.66‐, and 0.63‐fold) were measured by Western blotting. N=4. The right panel was the quantity of the Western blot. B, AFs were incubated with IL‐11 neutralizing antibody 1 hour before Ang II 24‐hour treatment. The mRNA levels of COL1a1 (1.00‐, 1.09‐, 1.40‐, and 1.52‐fold), ACTA2 (1.00‐, 0.91‐, 1.80‐, and 1.58‐fold), and KLF15 (1.00‐, 0.98‐, 0.37‐, and 0.51‐fold) were measured by quantitative reverse transcription–polymerase chain reaction. N=5. C, AFs were incubated with ribosomal S6 kinase inhibitor (CMK) (10 µmol/L) or ERK1/2 inhibitor (ERKi) (10 µmol/L) 1 hour before recombination mouse IL‐11 ( rmIL‐11) (5 ng/mL) 24‐hour treatment. The protein levels of phosphorylated p90RSK (p‐p90RSK) (1.00‐, 2.10‐, 0.93‐, and 0.74‐fold), p90RSK, phosphorylated ERK1/2 (p‐ERK1/2) (1.00‐, 10.71‐, 10.89‐, and 2.53‐fold), and ERK1/2 were measured by Western blotting. N=4. The right panel was the quantity of the Western blot. D, AFs were incubated with dimethyl sulfoxide, CMK (10 µmol/L), or ERKi (10 µmol/L) 1 hour before PBS or rmIL‐11 (5 ng/mL) 24‐hour treatment. The protein levels of COL1a1 (1.00‐, 1.40‐, 0.68‐, and 0.79‐fold) and ACTA2 (1.00‐, 1.13‐, 0.89‐, and 0.95‐fold) were measured by Western blotting. N=4. The right panel was the quantity of the Western blot. CON indicates control. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: For Ang II+IL‐11 neutralizing antibody group, wild‐type (WT) mice were treated with 1 mg/kg intravenous IL‐11 neutralizing antibody (R&D; MAB418) at 7, 9, 11, and 13 days after Ang II infusion.

Techniques: Incubation, Western Blot, Reverse Transcription Polymerase Chain Reaction