Journal: bioRxiv

Article Title: A novel mouse model of hypertensive emergency with multiorgan microvascular disease implicating the VEGFA/sFlt-1 balance

doi: 10.64898/2026.03.03.709451

Figure Lengend Snippet: A- Mice survival after hypertension onset (days after beginning of Ang II infusion). N= 20 C57BL6/J (B6J) and 129Sv (129) B- Mean daytime telemetry blood pressure N= 4 B6J and 129Sv. Values represent mean ± SEM. C- Mean nighttime telemetry blood pressure. N= 4 B6J and 129Sv. Values represent mean ± SEM.

Article Snippet: The hypertensive model was induced by subcutaneous infusion of angiotensin II (Ang II) (Alomone, GPA-100) at 1 μg/kg/min for 7 or 14 days, via osmotic minipumps (Alzet, model 1007D or 1002) in 10- to 16-week-old males.

Techniques:

Journal: bioRxiv

Article Title: A novel mouse model of hypertensive emergency with multiorgan microvascular disease implicating the VEGFA/sFlt-1 balance

doi: 10.64898/2026.03.03.709451

Figure Lengend Snippet: A- In vivo imaging (microm) performed on 129Sv (129) and C57BL6/J (B6J) mice 8 days after angiotensin-2 pump implantation shows haemorrhage (arrows) and hard exudates (asterisks). Haemorrhagic spots are only found on the 129Sv eye fundus and density of hard exudates is increased in 129 mice compared to B6J mice. B-Haemorrhagic spots (arrow heads) in retinas at day 7 of hypertensive challenge and associated quantification. Values represent mean ± SEM of 5 mice per group. * p < 0.05. C- Subretinal swelling assessed by OCT at day 7 of hypertensive challenge and associated quantification. The white arrow shows retinal detachment. D- Retinal capillary density assessed at day 7 of hypertensive challenge. Representative images of the retina vascular network stained with lectin antibody. Values represent mean ± SEM of 4 B6J mice and 8 129 mice. *** p < 0.001.

Article Snippet: The hypertensive model was induced by subcutaneous infusion of angiotensin II (Ang II) (Alomone, GPA-100) at 1 μg/kg/min for 7 or 14 days, via osmotic minipumps (Alzet, model 1007D or 1002) in 10- to 16-week-old males.

Techniques: In Vivo Imaging, Staining

Journal: bioRxiv

Article Title: A novel mouse model of hypertensive emergency with multiorgan microvascular disease implicating the VEGFA/sFlt-1 balance

doi: 10.64898/2026.03.03.709451

Figure Lengend Snippet: Typical ECG changes seen in SV129 mice treated for 2 weeks with angiotensin II (n = 5-7 mice/group. A. episode of atrial flutter B. atrial fibrillation. C. ventricular ectopy seen in the Ang II group; D. short run of ventricular tachycardia; E. sustained ventricular tachycardia degenerating in ventricular fibrillation and cardiac death.

Article Snippet: The hypertensive model was induced by subcutaneous infusion of angiotensin II (Ang II) (Alomone, GPA-100) at 1 μg/kg/min for 7 or 14 days, via osmotic minipumps (Alzet, model 1007D or 1002) in 10- to 16-week-old males.

Techniques:

Journal: bioRxiv

Article Title: Angiotensin II- Angiotensin II Receptor Type 1 Signaling Facilitates Gastric Cancer Metastasis via Kruppel-like Factor 4 Suppression and Tight Junction Breakdown

doi: 10.64898/2026.01.08.698396

Figure Lengend Snippet: (A) Representative IHC staining shows that the expression of ATII and AT1R is significantly elevated in GC tissues compared with adjacent normal gastric tissues. Original magnification, x20 (B) The violin plot shows that the IHC score for ATII and AT1R is significantly higher in non-metastatic and metastatic GC tissues than in adjacent normal gastric tissues. Significance was determined by 1-way ANOVA ( n = 64). ( C ) TCGA data show the expression of AGT and a stage-wise increase in expression of AGT in STAD. ( D) Kaplan-Meier survival curves from TCGA show OS and DFS of STAD patients grouped by AGT and AGTR1 expressions. High AGT expression is associated with significantly poor OS ( p = 0.02), and high AGTR1 expression is associated with significantly poor DFS ( p = 0.0035). GC, gastric cancer; IHC, immunohistochemistry; STAD, stomach adenocarcinoma; OS, overall survival; DFS, disease-free survival; TCGA, the cancer genome atlas; AGT, Angiotensin II; AGTR1, Angiotensin II receptor 1 . **P < 0.01 ; ****P < 0.0001. ns: no significance.

Article Snippet: The antibodies ATII (Cat# NB100-62346 Novus Biologicals, CO), AT1R (Cat# ab124505 Abcam, MA), KLF4 (Cat# ab216968 Abcam, MA and Cat# 4038S Cell Signaling Technology, Danvers, MA), claudin 1 (Cat# H00009076-M01 Abnova, Taipei, Taiwan), claudin 3 (Cat# NBP2-46299 Novus Biologicals, CO), claudin 4 (Cat# ab53156, Abcam, MA), Zonula occludens 1 (Cat# NBP2-80141 Novus Biologicals, CO), GAPDH (Cat#2118 Cell Signaling Technology, MA), HDAC1 (Cat# NB100-56340 Novus Biologicals, CO) were used for western blotting ( ).

Techniques: Immunohistochemistry, Expressing

Journal: bioRxiv

Article Title: Angiotensin II- Angiotensin II Receptor Type 1 Signaling Facilitates Gastric Cancer Metastasis via Kruppel-like Factor 4 Suppression and Tight Junction Breakdown

doi: 10.64898/2026.01.08.698396

Figure Lengend Snippet: (A) Enzymatic Immuno Assay (EIA), western blot, and qRT-PCR reveal expression of ATII in GC cells, MKN45, MKN7, AGS, and NCI-N87. For quantification, the mRNA level of the ATII gene was normalized to the housekeeping gene GAPDH. Data represented as mean±SD of three independent experiments. (B) Western blot and qRT-PCR data show the expression of AT1R in GC cells. For quantification, the mRNA level of the ATIR gene was normalized to the housekeeping gene GAPDH. Data represented as mean±SD of three independent experiments. Blockade of AT1R by the AT1R antagonist losartan (1µM) significantly reduces GC cell (C) proliferation, (D) migration (Original magnification, ×4), and (E) invasion (Original magnification, ×20). For the proliferation the data represented as mean±SEM and analyzed using 2-tailed t test ( n = 5). The percentage of wound closure is shown in bar diagrams. Data represented as mean±SD and analyzed using 2-tailed t test ( n = 3). Transwell assay shows that losartan treatment significantly reduces the invasive ability of MKN45 and NCI-N87 GC cells by inhibition of the AT1R receptors expressed in these cells. The number of invaded cells through the membrane in the control and losartan-treated groups is represented as a bar diagram. Data represented as mean±SEM and analyzed using 2-tailed t test ( n = 9). ( F) A volcano plot illustrating the differentially expressed genes (DEGs) obtained from RNA-seq analysis of losartan-treated MKN45 GC cancer cells compared to control cells. Losartan-treated GC cells with upregulated and downregulated genes are shown in red and blue colors respectively. ( G) The heatmap illustrates the differential expression of TJ-related genes in losartan-treated MKN45 cells compared to control cells. ( H) GSEA plot demonstrates a positive enrichment of the KEGG TJ gene set in losartan-treated MKN45 cells, suggesting that TJs are more intact upon treatment with losartan. I , Violin plot depicts the overall expression of TJ-related genes in losartan-treated MKN45 cells compared with control cells. *** P < 0.001, **** P < 0.0001. GC, gastric cancer; EIA, enzymatic immunoassay; GSEA, gene set enrichment analysis; KEGG, Kyoto encyclopedia of genes and genomes.

Article Snippet: The antibodies ATII (Cat# NB100-62346 Novus Biologicals, CO), AT1R (Cat# ab124505 Abcam, MA), KLF4 (Cat# ab216968 Abcam, MA and Cat# 4038S Cell Signaling Technology, Danvers, MA), claudin 1 (Cat# H00009076-M01 Abnova, Taipei, Taiwan), claudin 3 (Cat# NBP2-46299 Novus Biologicals, CO), claudin 4 (Cat# ab53156, Abcam, MA), Zonula occludens 1 (Cat# NBP2-80141 Novus Biologicals, CO), GAPDH (Cat#2118 Cell Signaling Technology, MA), HDAC1 (Cat# NB100-56340 Novus Biologicals, CO) were used for western blotting ( ).

Techniques: Immuno Assay, Western Blot, Quantitative RT-PCR, Expressing, Migration, Transwell Assay, Inhibition, Membrane, Control, RNA Sequencing, Quantitative Proteomics, Enzyme Immunoassay

Journal: bioRxiv

Article Title: Angiotensin II- Angiotensin II Receptor Type 1 Signaling Facilitates Gastric Cancer Metastasis via Kruppel-like Factor 4 Suppression and Tight Junction Breakdown

doi: 10.64898/2026.01.08.698396

Figure Lengend Snippet: (A) IHC staining shows that the expression of TJ proteins Claudin 1, 3, 4, and Zonula occludens 1 is markedly decreased in human GC tissues compared to adjacent normal gastric tissues. The IHC score was analyzed and shown by the dot plot comparing IHC scores for Claudin 1 ( n = 26 for NS; n = 27 for GC), 3 ( n = 19 for NS; n = 19 for GC), 4 ( n = 21 for NS; n = 19 for GC), and Zonula occludens 1 ( n = 22 for NS; n = 21 for GC) in two groups: NS and GC. Original magnification, ×20. The IHC score for each TJ protein is significantly higher in normal stomach tissues compared to GC tissues. Each dot represents an individual data point, and the horizontal lines represent the median for each group. Data analyzed using 2-tailed t test. ( B) Expression of ATII in human GC tissues shows an inverse correlation with the expression of TJ proteins, Claudin 1 ( n = 40), 3 ( n = 40), 4 ( n = 43), and Zonula occludens 1 ( n = 44). Correlation analysis between ATII expression and Claudins was performed using Pearson’s correlation test and the values are displayed on the graph. ( C) Expression of AT1R in human GC tissues shows an inverse correlation with the expression of TJ proteins, Claudin 1 ( n = 40), 3 ( n = 40), 4 ( n = 43), and Zonula occludens 1 ( n = 44). Correlation analysis between ATIR expression and Claudins was performed using Pearson’s correlation test and the values are displayed on the graph. ( D) IHC images and quantification of KLF4-positive cells indicate a gradual loss of KLF4 expression in GC tissues with disease progression. Original magnification, ×20. Significance was determined by 1-way ANOVA ( n = 64). (E) Quantitative analysis reveals a strong positive correlation between the percentage of KLF4-positive cells and the expression of these TJ proteins, Claudin 1 ( n = 40), 3 ( n = 40), 4 ( n = 43), and Zonula occludens 1 ( n = 43) in GC tissues. Correlation analysis between KLF4 expression and Claudins was performed using Pearson’s correlation test and the values are displayed on the graph. ** P < 0.01;*** P < 0.001; **** P < 0.0001. ns : no significance. GC, gastric cancer; TJ, tight junction; NS, normal stomach; IHC, immunohistochemistry.

Article Snippet: The antibodies ATII (Cat# NB100-62346 Novus Biologicals, CO), AT1R (Cat# ab124505 Abcam, MA), KLF4 (Cat# ab216968 Abcam, MA and Cat# 4038S Cell Signaling Technology, Danvers, MA), claudin 1 (Cat# H00009076-M01 Abnova, Taipei, Taiwan), claudin 3 (Cat# NBP2-46299 Novus Biologicals, CO), claudin 4 (Cat# ab53156, Abcam, MA), Zonula occludens 1 (Cat# NBP2-80141 Novus Biologicals, CO), GAPDH (Cat#2118 Cell Signaling Technology, MA), HDAC1 (Cat# NB100-56340 Novus Biologicals, CO) were used for western blotting ( ).

Techniques: Immunohistochemistry, Expressing, Biomarker Discovery

Journal: bioRxiv

Article Title: Angiotensin II- Angiotensin II Receptor Type 1 Signaling Facilitates Gastric Cancer Metastasis via Kruppel-like Factor 4 Suppression and Tight Junction Breakdown

doi: 10.64898/2026.01.08.698396

Figure Lengend Snippet: (A) The quantitative correlation analysis shows a significant negative correlation between the average IHC score for ATII/AT1R expression and the number of KLF4-positive cells. Correlation was performed using Pearson’s correlation test and the values are displayed on the graph ( n = 46). (B) cBioPortal data show that elevated mRNA expression of AGT and AGTR1 is negatively correlated with KLF4 expression in GC. *P < 0.05 ; **P < 0.01 ; ***P < 0.001 ; ****P < 0.0001. GC, gastric cancer; TJ, tight junction.

Article Snippet: The antibodies ATII (Cat# NB100-62346 Novus Biologicals, CO), AT1R (Cat# ab124505 Abcam, MA), KLF4 (Cat# ab216968 Abcam, MA and Cat# 4038S Cell Signaling Technology, Danvers, MA), claudin 1 (Cat# H00009076-M01 Abnova, Taipei, Taiwan), claudin 3 (Cat# NBP2-46299 Novus Biologicals, CO), claudin 4 (Cat# ab53156, Abcam, MA), Zonula occludens 1 (Cat# NBP2-80141 Novus Biologicals, CO), GAPDH (Cat#2118 Cell Signaling Technology, MA), HDAC1 (Cat# NB100-56340 Novus Biologicals, CO) were used for western blotting ( ).

Techniques: Expressing

Journal: bioRxiv

Article Title: Angiotensin II- Angiotensin II Receptor Type 1 Signaling Facilitates Gastric Cancer Metastasis via Kruppel-like Factor 4 Suppression and Tight Junction Breakdown

doi: 10.64898/2026.01.08.698396

Figure Lengend Snippet: (A) Schematic representation of MKN45 and NCI-N87 GC orthotopic implantations followed by treatment with AT1R blockers in MKN45 and NCI-N87 tumor-bearing athymic mice (created by BioRender). ( B) Mice transplanted with MKN45 and NCI-N87 cells show visible liver metastatic nodules. ( C) Orthotopic GC tumors show high expression of ATII and AT1R ( n = 11 per group). Original magnification, x20. ( D) Immunohistochemistry shows a significant change in expression of KLF4 in MKN45 and NCI-N87 orthotopic tumors, which are highly metastatic to the liver. Strong KLF4 expression is observed in normal mouse stomach tissues ( n = 11). Original magnification, x20. (E) In vivo administration of AT1R blockers, losartan and candesartan, for 28 consecutive days significantly reduces tumor growth in MKN45 and NCI-N87 orthotopic xenograft models, as measured by the wet tumor weight. Each dot represents a mouse. Data represented as mean±SEM. Significance was determined by 1-way ANOVA with Dunnett’s multiple comparisons test. (F) Treatment with losartan and candesartan also results in a significant reduction in distant metastasis. (G) qRT-PCR analysis shows significant upregulation of KLF4, CLDN1, CLDN3, CLDN4, and TJP1 transcripts in GC tissues from losartan-treated mice compared to those from untreated mice. For the quantification of mRNA levels, genes were normalized to the housekeeping gene GAPDH. Data represented as mean±SD and analyzed using 1-way ANOVA with Turkey’s multiple comparisons test (n = 3). ( H) Western blot analysis further confirms the significant increase in the expression of KLF4, Claudin 1 and Claudin 4 in mouse GC tissues upon losartan treatment. *P < 0.05 ; **P < 0.01 ; ***P < 0.001 ; ****P < 0.0001. GC, gastric cancer; IHC, immunohistochemistry; TJ, tight junction.

Article Snippet: The antibodies ATII (Cat# NB100-62346 Novus Biologicals, CO), AT1R (Cat# ab124505 Abcam, MA), KLF4 (Cat# ab216968 Abcam, MA and Cat# 4038S Cell Signaling Technology, Danvers, MA), claudin 1 (Cat# H00009076-M01 Abnova, Taipei, Taiwan), claudin 3 (Cat# NBP2-46299 Novus Biologicals, CO), claudin 4 (Cat# ab53156, Abcam, MA), Zonula occludens 1 (Cat# NBP2-80141 Novus Biologicals, CO), GAPDH (Cat#2118 Cell Signaling Technology, MA), HDAC1 (Cat# NB100-56340 Novus Biologicals, CO) were used for western blotting ( ).

Techniques: Expressing, Immunohistochemistry, In Vivo, Quantitative RT-PCR, Western Blot

Journal: Molecular Nutrition & Food Research

Article Title: Chlorogenic Acid Ameliorates Chronic Unpredictable Stress‐Induced Diminished Ovarian Reserve Through Ovarian Renin‐Angiotensin System

doi: 10.1002/mnfr.202400814

Figure Lengend Snippet: CGA improved DOR by reducing oxidative stress and apoptosis levels in mouse ovaries through its effects on the ACE‐AngII‐AT1R axis. (A) and (B) The qRT‐PCR analysis of the OVRAS components Ace , Agt , Agtr1a , Agtr1b , Ace2 , Agtr2 , and MasR ( n = 3). (C) Expression levels of oxidative stress and apoptosis molecular markers ( Bax , Bcl‐2 , Sod2 , and Gpx4 ) in the ovaries were detected by qRT‐PCR ( n = 3). (D–F) Quantification of ACE, AGT, AGTR1, ACE2, AGTR2, MasR, BAX, Bcl‐2, SOD2, and GPX4 protein expression. (G) and (H) Photographs of western blot showing the protein level of ACE, ACE2, AGT, AGTR1, AGTR2, MasR, BAX, Bcl‐2, GPX4, and SOD2. (I) and (J) Analysis of AngII and Ang(1‐7) in mouse ovary tissue homogenates collected from each group ( n = 3). CAT (K), GSH (L), and MDA (M) levels as oxidative stress indicators were determined in mouse ovary tissue homogenates ( n = 6). Results are expressed as the mean ± SD and one way ANOVA was used for between‐group comparisons, n represents the number of biological replicates. * p < 0.05, ** p < 0.01, *** p < 0.001, ns: not significant.

Article Snippet: Proteins were separated on SDS‐PAGE gels (Epizyme, China), transferred to PVDF membranes (Millipore, USA), and then blocked with 5% non‐fat milk at room temperature for 2 h. The following primary antibodies were used: AGT (Affinity, China, DF7976, 1:1000), AGTR1 (BOSTER, China, BM4949, 1:1000), AGTR2 (BOSTER, China, BM4557, 1:1000), MasR (Affinity, China, DF2818, 1:1000), ACE (Affinity, China, AF5197, 1:1000), ACE2 (Affinity, China, AF5165, 1:1000), BAX (Bimake, USA, A5131, 1:1000), Bcl‐2 (Affinity, China, BF9103, 1:1000), SOD2 (Selleck, USA, A5377, 1:1000), GPX4 (Selleck, USA, A5569, 1:1000), GAPDH (Proteintech, China, 60004‐1‐Ig, 1:10000).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Molecular Nutrition & Food Research

Article Title: Chlorogenic Acid Ameliorates Chronic Unpredictable Stress‐Induced Diminished Ovarian Reserve Through Ovarian Renin‐Angiotensin System

doi: 10.1002/mnfr.202400814

Figure Lengend Snippet: CGA decreased activity of ACE‐AngII‐AT1R axis as well as reduced oxidative stress and apoptosis levels of Dex‐induced KGN cells. (A) and (B) The qRT‐PCR analysis of OVRAS components ACE , AGT , AGTR1 , ACE2 , AGTR2 , and MasR mRNA expression level ( n = 3). (C) Expression levels of oxidative stress and apoptosis molecular markers ( BAX , Bcl‐2 , SOD2 , and GPX4 ) detected by qRT‐PCR ( n = 3). (G) and (H) The OVRAS, oxidative stress and apoptosis‐related protein expression in PVDF membrane. (D) and (E) Quantification of ACE, AGT, AGTR1, ACE2, AGTR2, and MasR protein expression ( n = 3). (H) Quantification of BAX, Bcl‐2, SOD2, and GPX4 protein expression ( n = 3). Results are expressed as the mean ± SD and one way ANOVA was used for between‐group comparisons, n represents the number of biological replicates. * p < 0.05, ** p < 0.01, *** p < 0.001, ns: not significant.

Article Snippet: Proteins were separated on SDS‐PAGE gels (Epizyme, China), transferred to PVDF membranes (Millipore, USA), and then blocked with 5% non‐fat milk at room temperature for 2 h. The following primary antibodies were used: AGT (Affinity, China, DF7976, 1:1000), AGTR1 (BOSTER, China, BM4949, 1:1000), AGTR2 (BOSTER, China, BM4557, 1:1000), MasR (Affinity, China, DF2818, 1:1000), ACE (Affinity, China, AF5197, 1:1000), ACE2 (Affinity, China, AF5165, 1:1000), BAX (Bimake, USA, A5131, 1:1000), Bcl‐2 (Affinity, China, BF9103, 1:1000), SOD2 (Selleck, USA, A5377, 1:1000), GPX4 (Selleck, USA, A5569, 1:1000), GAPDH (Proteintech, China, 60004‐1‐Ig, 1:10000).

Techniques: Activity Assay, Quantitative RT-PCR, Expressing, Membrane

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Activation of angiogenin expression in macrophages by lipopolysaccharide via the TLR4/NF-κB pathway in colitis

doi: 10.3724/abbs.2024013

Figure Lengend Snippet: Table 1 Sequences of primers and siRNAs used in this study

Article Snippet: The membrane was blocked with 5% nonfat milk and then incubated with primary antibodies, including anti-ANG antibody (prepared in our own laboratory) and anti-ACTB antibody (#81115-1-RR; Proteintech, Chicago, USA), in TBST (Tris-buffered saline, 0.1% Tween 20) buffer at 4°C overnight.

Techniques: Negative Control

Journal: Scientific Reports

Article Title: Sniffer cells for the detection of neural Angiotensin II in vitro

doi: 10.1038/s41598-019-45262-4

Figure Lengend Snippet: ANG II mediated increases in sniffer cell fluorescence. ( A ) ANG II (100 nM) induced a robust but transient increase in GCaMP fluorescence that was blocked by the AT1aR receptor antagonist Losartan (10 µM). Control n = 39, Losartan n = 38. ( B ) Data shows that bath application of glutamate (50 µM), GABA (50 µM), and carbachol (50 µM) failed to change fluorescent intensity of sniffer cells transfected with GCaMP (n = 29) or GCaMP + AT1aR (n = 38). ANG II (100 nM) did increase fluorescent intensity of sniffer cells, but only in sniffer cells transfected with GCaMP + AT1aR. Dose-dependent effects of ANG II and related compounds were also measured. ( C ) GCaMP + AT1aR sniffer cells exhibit dose-dependent increases in fluorescence in response to bath application of ANG II (100 nM, n = 10). ( D ) Bath application of ANG III induced a dose-dependent increase in GCaMP + AT1aR sniffer cell fluorescence. Bath application of ANG (1–7) or bradykinin did not induce a change in GCaMP + AT1aR sniffer cell fluorescence at any of the doses tested (0.1–100 nM, n = 17–41). ( E ) R-GECO + AT1aR sniffer cells exhibit dose-dependent increases in fluorescence in response to bath application of ANG II (n = 17). R-GECO only cells did not respond to ANG II (n = 27). ( F ) ANG II-mediated increases in R-GECO + AT1aR are blocked by bath application of Losartan (10 µM, n = 17). *p < 0.05, **p < 0.01.

Article Snippet: Carbachol (50 µM), Angiotensin 1–7 (0.1–100 nM), Bradykinin (0.1–100 nM), and Losartan (10 µM) were purchased from Tocris (Minneapolis, MN) and Angiotensin III (0.1–100 nM) was purchased from Alomone Labs (Jerusalem, Israel).

Techniques: Fluorescence, Transfection