Journal: Molecular Medicine

Article Title: NAT10 induces N4-acetylcytidine modification of AdipoR1-mediated mitochondrial biogenesis against endothelial-to-mesenchymal transition in hypertension

doi: 10.1186/s10020-025-01321-3

Figure Lengend Snippet: High NAT10 and ac4C levels in hypertension groups compared to the control groups. WB assay and the quantitative analysis of NAT10 level in hypertensive mice descending thoracic aortic tissues ( A , B ; n = 3 ) , SHRs descending thoracic aortic samples ( A , C ; n = 3 ) and Ang II treated HUVECs ( A , D ; n = 3 ) . ( E) The ac4C level in hypertensive mice descending thoracic aortic tissues, SHRs descending thoracic aortic samples and Ang II treated HUVECs ( n = 3). ( F) Representative IF staining of NAT10 and CD31 in hypertensive mice descending thoracic aortic tissues, SHRs descending thoracic aortic samples and the control tissues. Fluorescence in green represents CD31, while fluorescence in red represents NAT10 and fluorescence in blue represents DAPI. Scale bar, 100 μm. (representative images; n = 6). Data are presented as mean ± SD. ** p < 0.01. Statistical tests were performed using unpaired two-tailed Student’s t-test

Article Snippet: The descending thoracic aortic sections were blocked with blocking buffer (3% bovine serum albumin in PBS) for 60 min and stained with primary antibodies containing NAT10 (Abcam, ab194297, rabbit, 1:500), CD31 (MCE, YA806, mouse, 1:100) and SM22α (Proteintech, 10493-1-AP, rabbit, 1:100) overnight at 4 °C and the corresponding Alexa-conjugated secondary antibody containing Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 488) (Abcam, ab150105, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 594) (Abcam, ab150080, 1:1000), Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 594) (Abcam, ab150108, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) (Abcam, ab150077, 1:1000) at room temperature for 1 h. The nuclei were counterstained with DAPI (C0065, Solarbio) for 5 min, and the fluorescence images were captured under a fluorescence microscope (Olympus).

Techniques: Control, Staining, Fluorescence, Two Tailed Test

Journal: Molecular Medicine

Article Title: NAT10 induces N4-acetylcytidine modification of AdipoR1-mediated mitochondrial biogenesis against endothelial-to-mesenchymal transition in hypertension

doi: 10.1186/s10020-025-01321-3

Figure Lengend Snippet: NAT10 overexpression inhibited endothelial dysfunction and EndMT in hypertension. ( A , B ) WB assay and the quantitative analysis of NAT10 level in HUVECs after Ang II stimulation ( n = 3). ( C ) ECs proliferation analysis between OE-NAT10 and OE-NC group after Ang II stimulation ( n = 3). ( D ) ECs migration analysis between OE-NAT10 and OE-NC group after Ang II stimulation by Transwell migration assay ( n = 3). Scale bar, 100 μm. ( E ) ECs angiogenesis analysis between OE-NAT10 and OE-NC group after Ang II stimulation by tube formation assay ( n = 3). Scale bar, 100 μm. ( F , G ) WB assay and the quantitative analysis of CD31, VE-cadherin, SM22α and N-cadherin levels in HUVECs after Ang II stimulation ( n = 3). ( H ) H&E staining of descending thoracic aortic sections and the relative wall thickness of each group ( n = 6). Scale bar, 100 μm. ( I ) Masson staining of each group and quantitative analysis of the fibrotic area ( n = 6). Scale bar, 100 μm. ( J ) IF staining of CD31 and SM22α levels in OE-NAT10 and OE-NC group ( n = 6). Fluorescence in red represents CD31, while fluorescence in green represents SM22α and fluorescence in blue represents DAPI. Scale bar, 100 μm. ( K , L ) WB assay of CD31, VE-cadherin, SM22α and N-cadherin levels in OE-NAT10 and OE-NC group ( n = 6). Data represented as mean ± SD from three independent experiments. ** p < 0.01. Statistical tests were performed using unpaired two-tailed Student’s t-test (B-G, L) and Mann–Whitney U test (H, I)

Article Snippet: The descending thoracic aortic sections were blocked with blocking buffer (3% bovine serum albumin in PBS) for 60 min and stained with primary antibodies containing NAT10 (Abcam, ab194297, rabbit, 1:500), CD31 (MCE, YA806, mouse, 1:100) and SM22α (Proteintech, 10493-1-AP, rabbit, 1:100) overnight at 4 °C and the corresponding Alexa-conjugated secondary antibody containing Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 488) (Abcam, ab150105, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 594) (Abcam, ab150080, 1:1000), Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 594) (Abcam, ab150108, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) (Abcam, ab150077, 1:1000) at room temperature for 1 h. The nuclei were counterstained with DAPI (C0065, Solarbio) for 5 min, and the fluorescence images were captured under a fluorescence microscope (Olympus).

Techniques: Over Expression, Migration, Transwell Migration Assay, Tube Formation Assay, Staining, Fluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Molecular Medicine

Article Title: NAT10 induces N4-acetylcytidine modification of AdipoR1-mediated mitochondrial biogenesis against endothelial-to-mesenchymal transition in hypertension

doi: 10.1186/s10020-025-01321-3

Figure Lengend Snippet: NAT10 depletion induced endothelial dysfunction and EndMT in hypertension. (A, B) WB assay and the quantitative analysis of NAT10 level in HUVECs after Ang II stimulation ( n = 3). (C) ECs proliferation analysis between sh-NAT10 and sh-NC group after Ang II stimulation ( n = 3). (D) ECs migration analysis between sh-NAT10 and sh-NC group after Ang II stimulation ( n = 3). Scale bar, 100 μm. (E) ECs angiogenesis analysis between sh-NAT10 and sh-NC group after Ang II stimulation ( n = 3). Scale bar, 100 μm. (F, G) WB assay and the quantitative analysis of CD31, VE-cadherin, SM22α and N-cadherin levels in HUVECs after Ang II stimulation ( n = 3). (H) H&E staining of descending thoracic aortic sections and the relative wall thickness of each group ( n = 6). Scale bar, 100 μm. (I) Masson staining of each group and quantitative analysis of the fibrotic area ( n = 6). Scale bar, 100 μm. (J) IF staining of CD31 and SM22α levels in sh-NAT10 and sh-NC group ( n = 6). Fluorescence in red represents CD31, while fluorescence in green represents SM22α and fluorescence in blue represents DAPI. Scale bar, 100 μm. (K, L) WB assay of CD31, VE-cadherin, SM22α and N-cadherin levels in sh-NAT10 and sh-NC group ( n = 6). Data represented as mean ± SD from three independent experiments. ** p < 0.01. Statistical tests were performed using unpaired two-tailed Student’s t-test (B-G, L) and Mann–Whitney U test (H, I)

Article Snippet: The descending thoracic aortic sections were blocked with blocking buffer (3% bovine serum albumin in PBS) for 60 min and stained with primary antibodies containing NAT10 (Abcam, ab194297, rabbit, 1:500), CD31 (MCE, YA806, mouse, 1:100) and SM22α (Proteintech, 10493-1-AP, rabbit, 1:100) overnight at 4 °C and the corresponding Alexa-conjugated secondary antibody containing Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 488) (Abcam, ab150105, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 594) (Abcam, ab150080, 1:1000), Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 594) (Abcam, ab150108, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) (Abcam, ab150077, 1:1000) at room temperature for 1 h. The nuclei were counterstained with DAPI (C0065, Solarbio) for 5 min, and the fluorescence images were captured under a fluorescence microscope (Olympus).

Techniques: Migration, Staining, Fluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Molecular Medicine

Article Title: NAT10 induces N4-acetylcytidine modification of AdipoR1-mediated mitochondrial biogenesis against endothelial-to-mesenchymal transition in hypertension

doi: 10.1186/s10020-025-01321-3

Figure Lengend Snippet: Remodelin induced endothelial dysfunction and EndMT in hypertension. ( A , B) WB assay and the quantitative analysis of NAT10 level in HUVECs after Ang II stimulation ( n = 3). ( C - E) ECs proliferation, migration and angiogenesis analysis between remodelin and control group after Ang II stimulation ( n = 3). Scale bar, 100 μm. ( F , G ) WB assay and the quantitative analysis of CD31, VE-cadherin, SM22α and N-cadherin levels in HUVECs after Ang II stimulation ( n = 3). ( H ) H&E staining of descending thoracic aortic sections and the relative wall thickness of remodelin and control group ( n = 6). Scale bar, 50 μm. ( I) Masson staining of remodelin and control group and quantitative analysis of the fibrotic area ( n = 6). Scale bar, 50 μm. ( J) IF staining of CD31 and SM22α levels in remodelin and control group ( n = 6). Fluorescence in red represents CD31, while fluorescence in green represents SM22α and fluorescence in blue represents DAPI. Scale bar, 50 μm. ( K , L) WB assay of CD31, VE-cadherin, SM22α and N-cadherin levels in remodelin and control group ( n = 6). Data represented as mean ± SD from three independent experiments. ** p < 0.01. Statistical tests were performed using unpaired two-tailed Student’s t-test (B-G, L) and Mann–Whitney U test (H, I)

Article Snippet: The descending thoracic aortic sections were blocked with blocking buffer (3% bovine serum albumin in PBS) for 60 min and stained with primary antibodies containing NAT10 (Abcam, ab194297, rabbit, 1:500), CD31 (MCE, YA806, mouse, 1:100) and SM22α (Proteintech, 10493-1-AP, rabbit, 1:100) overnight at 4 °C and the corresponding Alexa-conjugated secondary antibody containing Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 488) (Abcam, ab150105, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 594) (Abcam, ab150080, 1:1000), Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 594) (Abcam, ab150108, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) (Abcam, ab150077, 1:1000) at room temperature for 1 h. The nuclei were counterstained with DAPI (C0065, Solarbio) for 5 min, and the fluorescence images were captured under a fluorescence microscope (Olympus).

Techniques: Migration, Control, Staining, Fluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Molecular Medicine

Article Title: NAT10 induces N4-acetylcytidine modification of AdipoR1-mediated mitochondrial biogenesis against endothelial-to-mesenchymal transition in hypertension

doi: 10.1186/s10020-025-01321-3

Figure Lengend Snippet: AdipoR1 is a downstream target of NAT10 in Ang II treated ECs. ( A , B) WB assay and the quantitative analysis of AdipoR1 level in HUVECs after Ang II stimulation ( n = 3). ( C) ECs proliferation analysis after Ang II stimulation ( n = 3). ( D) ECs migration analysis after Ang II stimulation ( n = 3). Scale bar, 100 μm. ( E ) ECs angiogenesis analysis after Ang II stimulation ( n = 3). Scale bar, 100 μm. ( F , G) WB assay and the quantitative analysis of CD31, VE-cadherin, SM22α and N-cadherin levels in HUVECs after Ang II stimulation ( n = 3). ( H) H&E staining of descending thoracic aortic sections and the relative wall thickness of each group ( n = 6). Scale bar, 100 μm. ( I) Masson staining of each group and quantitative analysis of the fibrotic area ( n = 6). Scale bar, 100 μm. ( J) IF staining of CD31 and SM22α levels in each group ( n = 6). Fluorescence in red represents CD31, while fluorescence in green represents SM22α and fluorescence in blue represents DAPI. Scale bar, 100 μm. ( K , L) WB assay of CD31, VE-cadherin, SM22α and N-cadherin levels in each group ( n = 6). Data represented as mean ± SD from three independent experiments. ** p < 0.01, # p < 0.05, ## p < 0.01. Statistical analysis was performed using t-test (unpaired, two-sided) between two groups or one-way ANOVA followed by Tukey’s multiple comparisons test for multiple-group comparisons

Article Snippet: The descending thoracic aortic sections were blocked with blocking buffer (3% bovine serum albumin in PBS) for 60 min and stained with primary antibodies containing NAT10 (Abcam, ab194297, rabbit, 1:500), CD31 (MCE, YA806, mouse, 1:100) and SM22α (Proteintech, 10493-1-AP, rabbit, 1:100) overnight at 4 °C and the corresponding Alexa-conjugated secondary antibody containing Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 488) (Abcam, ab150105, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 594) (Abcam, ab150080, 1:1000), Donkey Anti-Mouse IgG H&L (Alexa Fluor ® 594) (Abcam, ab150108, 1:500), Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) (Abcam, ab150077, 1:1000) at room temperature for 1 h. The nuclei were counterstained with DAPI (C0065, Solarbio) for 5 min, and the fluorescence images were captured under a fluorescence microscope (Olympus).

Techniques: Migration, Staining, Fluorescence

Journal: iScience

Article Title: In vitro vascularized immunocompetent patient-derived model to test cancer therapies

doi: 10.1016/j.isci.2023.108094

Figure Lengend Snippet:

Article Snippet: Mouse anti-CD31 , Cell Applications , Cat# CB13678.

Techniques: Plasmid Preparation, Recombinant, Lysis, RNA Extraction, Amplification, DC Protein Assay, Enzyme-linked Immunosorbent Assay, Sequencing, Software, Fluorescence, Microscopy

Journal: Molecular therapy. Nucleic acids

Article Title: Icariin attenuates endothelial-mesenchymal transition via H19/miR-148b-3p/ELF5 in ox-LDL-stimulated HUVECs.

doi: 10.1016/j.omtn.2020.11.021

Figure Lengend Snippet: Figure 1. ICA suppressed ox-LDL-induced EndMT in HUVECs (A–E) Expression levels of CD31, VE-cadherin, a-SMA, and S100A4 in HUVECs exposed to 200 mM ox-LDL or ox-LDL+10 mM ICA, determined using western blotting. ##p < 0.01 versus control group; **p < 0.01 versus ox-LDL group; n = 3. Measurement data are expressed as mean ± SD. ICA, icariin; ox-LDL, oxidized low-density lipoprotein; EndMT, endothelial-mesenchymal transition; HUVEC, human umbilical vein endothelial cell.

Article Snippet: The proteins were then transferred onto polyvinylidene fluoride (PVDF) membranes and incubated with TBST containing 5% BSA for 1 h at 20 C–25 C. The PVDF membranes were probed overnight with CD31 (lot no. 3528S, Cell Signaling Technology, Danvers, MA, USA), VE-cadherin Molecular Therapy: Nucleic Acids Vol.

Techniques: Expressing, Western Blot, Control

Journal: Molecular therapy. Nucleic acids

Article Title: Icariin attenuates endothelial-mesenchymal transition via H19/miR-148b-3p/ELF5 in ox-LDL-stimulated HUVECs.

doi: 10.1016/j.omtn.2020.11.021

Figure Lengend Snippet: Figure 2. ICA promoted ox-LDL-induced H19 expression, and H19 affected EndMT in HUVECs (A) The expression of H19 in HUVECs in control, ox-LDL, or ox-LDL+ICA groups was detected using quantitative real-time PCR. (B) Following transfection with recombinant H19 plasmids or sh-H19, the expression of H19 was detected using quantitative real-time PCR. (C–G) The expression of CD31, VE-cadherin, a-SMA, and S100A4 in ox-LDL- treated HUVECs was determined using western blot analysis. (H) Transwell assays were used to assess the migration capacity of HUVECs incubated for 24 h. (I) HUVECs were incubated for 24 h and subjected to a wound-healing assay to assess cell migration. Images were acquired at 0 and 24 h. *p < 0.05, **p < 0.01 versus pLVX-negative control (NC) group; #p < 0.05, ##p < 0.01 versus sh-NC group; n = 3. Measurement data are expressed as mean ± SD. Scale bars, 100 mm.

Article Snippet: The proteins were then transferred onto polyvinylidene fluoride (PVDF) membranes and incubated with TBST containing 5% BSA for 1 h at 20 C–25 C. The PVDF membranes were probed overnight with CD31 (lot no. 3528S, Cell Signaling Technology, Danvers, MA, USA), VE-cadherin Molecular Therapy: Nucleic Acids Vol.

Techniques: Expressing, Control, Real-time Polymerase Chain Reaction, Transfection, Recombinant, Western Blot, Migration, Incubation, Wound Healing Assay, Negative Control

Journal: Molecular therapy. Nucleic acids

Article Title: Icariin attenuates endothelial-mesenchymal transition via H19/miR-148b-3p/ELF5 in ox-LDL-stimulated HUVECs.

doi: 10.1016/j.omtn.2020.11.021

Figure Lengend Snippet: Figure 3. H19 knockdown attenuated the inhibitory effect of ICA on EndMT (A–E) Western blot analysis was performed to detect the expression of CD31, VE-cadherin, a-SMA, and S100A4 in HUVECs. (F and G) Transwell (F) and wound-healing assays (G) were used to assess cell migration in HUVECs treated with ox-LDL with or without ICA for 24 h. #p < 0.05, ##p < 0.01 versus control group; *p < 0.05, **p < 0.01 versus ox-LDL group; Dp < 0.05, DDp < 0.01 versus ox-LDL+ICA+sh-NC group; n = 3. Measurement data are expressed as mean ± SD. Scale bars, 100 mm.

Article Snippet: The proteins were then transferred onto polyvinylidene fluoride (PVDF) membranes and incubated with TBST containing 5% BSA for 1 h at 20 C–25 C. The PVDF membranes were probed overnight with CD31 (lot no. 3528S, Cell Signaling Technology, Danvers, MA, USA), VE-cadherin Molecular Therapy: Nucleic Acids Vol.

Techniques: Knockdown, Western Blot, Expressing, Migration, Control

Journal: Molecular therapy. Nucleic acids

Article Title: Icariin attenuates endothelial-mesenchymal transition via H19/miR-148b-3p/ELF5 in ox-LDL-stimulated HUVECs.

doi: 10.1016/j.omtn.2020.11.021

Figure Lengend Snippet: Figure 6. miR-148b-3p overexpression reversed H19-mitigated EndMT in ox-LDL-treated HUVECs Cells were transfected with H19 plasmids or miR-148b-3p mimic and exposed to ox-LDL. (A–E) After that, the expression levels of CD31, VE-cadherin, a-SMA, and S100A4 in HUVECs were determined using western blot analysis. (F and G) Cell migration capacity was measured using wound-healing (F) and transwell assays (G). *p < 0.05, **p < 0.01 versus pLVX-NC+miR-NC group; #p < 0.05, ##p < 0.01 versus pLVX-H19+miR-NC group; n = 3. Measurement data are expressed as mean ± SD. Scale bars, 100 mm.

Article Snippet: The proteins were then transferred onto polyvinylidene fluoride (PVDF) membranes and incubated with TBST containing 5% BSA for 1 h at 20 C–25 C. The PVDF membranes were probed overnight with CD31 (lot no. 3528S, Cell Signaling Technology, Danvers, MA, USA), VE-cadherin Molecular Therapy: Nucleic Acids Vol.

Techniques: Over Expression, Transfection, Expressing, Western Blot, Migration

Journal: Molecular therapy. Nucleic acids

Article Title: Icariin attenuates endothelial-mesenchymal transition via H19/miR-148b-3p/ELF5 in ox-LDL-stimulated HUVECs.

doi: 10.1016/j.omtn.2020.11.021

Figure Lengend Snippet: Figure 8. ICA inhibited the expression of TGF-b through H19, and galunisertib reversed the promoting effect of sh-H19 on EndMT (A) Cells were transfected with sh-H19 and exposed to ox-LDL (with or without ICA); western blot analysis was performed to detect the expression of transforming growth factor-b (TGF-b) in HUVECs. (B–G) After adding galunisertib, the expression of TGF-b, CD31, VE-cadherin, a-SMA, and S100A4 in HUVECs was also determined using western blot analysis. #p < 0.05, ##p < 0.01 versus control group; *p < 0.05, **p < 0.01 versus ox-LDL group; Dp < 0.05, DDp < 0.01 versus ox-LDL+ICA group; Vp < 0.05, VVp < 0.01 versus ox-LDL+ICA+sh-H19 group; &p < 0.05, &&p < 0.01 versus ox-LDL+ICA+sh-NC group; n = 3. Measurement data are expressed as mean ± SD.

Article Snippet: The proteins were then transferred onto polyvinylidene fluoride (PVDF) membranes and incubated with TBST containing 5% BSA for 1 h at 20 C–25 C. The PVDF membranes were probed overnight with CD31 (lot no. 3528S, Cell Signaling Technology, Danvers, MA, USA), VE-cadherin Molecular Therapy: Nucleic Acids Vol.

Techniques: Expressing, Transfection, Western Blot, Control

Journal: International journal of cardiology

Article Title: Enhanced external counterpulsation inhibits endothelial apoptosis via modulation of BIRC2 and Apaf-1 genes in porcine hypercholesterolemia.

doi: 10.1016/j.ijcard.2013.11.033

Figure Lengend Snippet: Fig. 2. Isolation and identification of vascular endothelial cells from the aortic endothelium with collagenase. Representative photomicrographs (×400) of isolated cells treated histocytochemically with CD31 antibody and incubated with DiI-Ac-LDL. Cells with cytomembrane and cytoplasmic staining of amber color indicate vascular endothelial cells (black arrow), and cells with red fluorescence were identified as viable vascular endothelial cells (white arrow), which consisted of the majority of total treated cells, indicating a successful en- dothelial cell isolation from aortic endothelium.

Article Snippet: Then mouse polyclonal CD31 antibody (at 1:100 dilution; Boster Biological Technology, Inc, Wuhan, China) was applied and incubated at 37 °C for 1 h, then stained with SABC, enhanced by DAB, dehydrated in a graded ethanol series, and followed by dimethylbenzene treatment to improve transparency, and finally mounted with neutral gum.

Techniques: Isolation, Incubation, Staining, Cell Isolation

Journal: PloS one

Article Title: SIRT1 regulates endothelial Notch signaling in lung cancer.

doi: 10.1371/journal.pone.0045331

Figure Lengend Snippet: Figure 1. Isolation of lung cancer xenograft-derived ECs. LLC xenografts were resected from mice injected subcutaneously at the dorsal flank with LLC cells (36106 suspended in 50 mL PBS) for 30 days. After removing obvious necrotic tissues or extra fatty compositions, the minced tissues were ground on ice using a glass grinder and were then filtered through cell strainers to eliminate tissue debris. (A) The CD31-expressing lung cancer- derived ECs were isolated from the single-cell suspension by immunomagnetic sorting, as evidenced by scanning microscopy, and cultured in vitro. (B) CD31 was detected in the isolated lung cancer-derived ECs using immunofluorescence. CD31 antibody staining of the lung cancer-derived EC membranes is shown in red, and nuclear DAPI staining is shown in blue. (C) Total protein was isolated from enriched lung cancer-derived ECs, bEnd.3 cells (positive control), and MLE-12 (negative control) cells. Western blot analysis was performed using antibodies against eNOS and E-cadherin, and b-actin was used as the internal control. doi:10.1371/journal.pone.0045331.g001

Article Snippet: The lung cancer-derived ECs were eluted, and CD31 expression was detected using a CD31 monoclonal antibody (Cell Signaling Technology, USA) and staining with NorthernLights 557-conjugated anti-mouse IgG secondary antibody.

Techniques: Isolation, Derivative Assay, Injection, Expressing, Suspension, Microscopy, Cell Culture, In Vitro, Immunofluorescence, Staining, Positive Control, Negative Control, Western Blot, Control

Journal: PloS one

Article Title: SIRT1 regulates endothelial Notch signaling in lung cancer.

doi: 10.1371/journal.pone.0045331

Figure Lengend Snippet: Figure 4. Effect of SIRT1 on tumor neovascularization in LLC xenografts. (A) SIRT1 promotes angiogenesis in vivo. Matrigel plugs were subcutaneously injected into the abdomens of control or SIRT1-transgenic mice, and the plugs were extracted 7 days later to determine the extent of vascularization. The amount of hemoglobin present in the plugs was quantified as an indicator of the formation of functional blood vessels (mean 6 SD; n = 10 for each group). * P,0.05 as compared to the control. (B) Changes in the median tumor volumes measured in wild-type (control) C57BL/6J, SIRT1, or SIRT1 (H363Y) mice following inoculation with LLC cells for the indicated time period. * P,0.05 as compared to the SIRT1 mice. (C) Photomicrographs of CD31 IHC staining in sections of LLC xenografts (magnification, 6200) from wild-type, SIRT1 or SIRT1(H363Y) mice. When the xenograft tumor volumes reached approximately 100 mm3, the mice were randomly assigned to the control arm (n = 8) or the experimental arm

Article Snippet: The lung cancer-derived ECs were eluted, and CD31 expression was detected using a CD31 monoclonal antibody (Cell Signaling Technology, USA) and staining with NorthernLights 557-conjugated anti-mouse IgG secondary antibody.

Techniques: In Vivo, Injection, Control, Transgenic Assay, Functional Assay, Immunohistochemistry