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: Cancer Cell International

Article Title: Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression

doi: 10.1186/s12935-025-03841-8

Figure Lengend Snippet: Pyk2 expression was significantly elevated in identified LCSCs. (A-B) The surface marker proteins CD133 and EPCAM of HepG2 and HepG2-LCSCs were measured by flow cytometry. (C-D) The surface marker proteins CD133 and EPCAM of HCCLM3 and HCCLM3-LCSCs were measured by flow cytometry. (E-F) The expression of Pyk2 in HepG2, HCCLM3, and relative LCSCs were measured using WB. Data are presented as mean ± standard deviation; * P < 0.05, ** P < 0.01, *** P < 0.001; n = 3

Article Snippet: A hydrophobic pen was used to draw a circle around the tissue sections, and endogenous peroxidase activity was blocked, followed by overnight incubation with the primary antibody against Pyk2 (dilution 1:100, 17592-1-AP, Proteintech, Wuhan, China) and Src (dilution 1:50, #2105, Cell signaling technology, Massachusetts, USA) at 4 °C.

Techniques: Expressing, Marker, Flow Cytometry, Standard Deviation

Journal: Cancer Cell International

Article Title: Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression

doi: 10.1186/s12935-025-03841-8

Figure Lengend Snippet: Construction of Pyk2 knockdown LCSCs. (A-B) The Pyk2 knockdown LCSCs were constructed by siRNA transfection. (C-D) The mRNA levels of Pyk2 in HepG2-LCSCs siPyk2 and HCCLM3-LCSCs siPyk2 were determined by RT-qPCR. (E-F) The protein expressions of Pyk2 in HepG2-LCSCs siPyk2 and HCCLM3-LCSCs siPyk2 s were determined by WB. Data are presented as mean ± standard deviation; * P < 0.05, ** P < 0.01, *** P < 0.001; n = 3

Article Snippet: A hydrophobic pen was used to draw a circle around the tissue sections, and endogenous peroxidase activity was blocked, followed by overnight incubation with the primary antibody against Pyk2 (dilution 1:100, 17592-1-AP, Proteintech, Wuhan, China) and Src (dilution 1:50, #2105, Cell signaling technology, Massachusetts, USA) at 4 °C.

Techniques: Knockdown, Construct, Transfection, Quantitative RT-PCR, Standard Deviation

Journal: Cancer Cell International

Article Title: Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression

doi: 10.1186/s12935-025-03841-8

Figure Lengend Snippet: Pyk2 expression maintain the stemness of LCSCs. (A-B) Colony formation assays of HepG2-LCSCs (A) and HCCLM3-LCSCs (B) transfected with Pyk2 siRNA (siPyk2) or negative control (NC). (C-D) Tumorsphere formation assays in the same groups as above for HepG2-LCSCs (C) and HCCLM3-LCSCs (D) . (E-F) Wound healing assays showing migratory capacity at 0 h and 24 h in HepG2-LCSCs (E) and HCCLM3-LCSCs (F) after Pyk2 knockdown. (G-H) Transwell invasion assays evaluating invasive ability of HepG2-LCSCs (G) and HCCLM3-LCSCs (H) under Pyk2 knockdown versus control conditions. (I-J) RT-qPCR analysis of stemness-related gene expression (Nanog, Oct4, Sox2, KLF4, Bmi1) in HepG2-LCSCs (I) and HCCLM3-LCSCs (J) following siPyk2 or NC treatment. (K-L) Cell proliferation and cisplatin sensitivity assessed by CCK-8 assay in HepG2-LCSCs (K) and HCCLM3-LCSCs (L) , respectively. Data are presented as mean ± standard deviation; * P < 0.05, ** P < 0.01, *** P < 0.001; n = 3

Article Snippet: A hydrophobic pen was used to draw a circle around the tissue sections, and endogenous peroxidase activity was blocked, followed by overnight incubation with the primary antibody against Pyk2 (dilution 1:100, 17592-1-AP, Proteintech, Wuhan, China) and Src (dilution 1:50, #2105, Cell signaling technology, Massachusetts, USA) at 4 °C.

Techniques: Expressing, Transfection, Negative Control, Knockdown, Control, Quantitative RT-PCR, Gene Expression, CCK-8 Assay, Standard Deviation

Journal: Cancer Cell International

Article Title: Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression

doi: 10.1186/s12935-025-03841-8

Figure Lengend Snippet: miR-23b-3p inhibits Pyk2/Src signaling in LCSCs. (A) Dual-luciferase reporter assay in HCCLM3-LCSCs co-transfected with wild-type or mutant Pyk2 3’-UTR constructs and miR-23b-3p mimic or inhibitor, demonstrating direct binding between miR-23b-3p and the Pyk2 3’-UTR. (B) RT-qPCR quantification of endogenous miR-23b-3p expression in HepG2, HCCLM3, and HCCLM3-LCSC cells. (C) Pyk2 mRNA levels measured by RT-qPCR in HCCLM3-LCSCs after transfection with miR-23b-3p inhibitor. (D-F) Western blot and quantitative analysis of total Pyk2 (D) , phosphorylated Pyk2 (E) , and p-Pyk2/Pyk2 ratio (F) in HCCLM3-LCSCs transfected with siPyk2, or co-transfected with siPyk2 and miR-23b-3p inhibitor. (G-I) Western blot and quantification of total Src (G) , phosphorylated Src (H) , and p-Src/Src ratio (I) in the same treatment groups. Data are presented as mean ± standard deviation; * P < 0.05, ** P < 0.01, *** P < 0.001; n = 3

Article Snippet: A hydrophobic pen was used to draw a circle around the tissue sections, and endogenous peroxidase activity was blocked, followed by overnight incubation with the primary antibody against Pyk2 (dilution 1:100, 17592-1-AP, Proteintech, Wuhan, China) and Src (dilution 1:50, #2105, Cell signaling technology, Massachusetts, USA) at 4 °C.

Techniques: Luciferase, Reporter Assay, Transfection, Mutagenesis, Construct, Binding Assay, Quantitative RT-PCR, Expressing, Western Blot, Standard Deviation

Journal: Cancer Cell International

Article Title: Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression

doi: 10.1186/s12935-025-03841-8

Figure Lengend Snippet: miR-23b-3p down-regulates Pyk2 expression to reduce the proliferation, migration, invasion, and chemosensitivity of LCSCs. (A-B) Wound healing assays evaluating the migration rate of HCCLM3-LCSCs transfected with miR-23b-3p inhibitor (A ), as well as siPyk2 and/or miR-23b-3p inhibitor (B) . (C-D) Transwell invasion assays assessing invasive ability of the same treatment groups as in (A-B) , showing increased invasion upon miR-23b-3p inhibition and partial rescue by siPyk2. (E-F) CCK-8 assays quantifying proliferation (E) and cisplatin-induced chemosensitivity (F) in HCCLM3-LCSCs after miR-23b-3p inhibition, with or without Pyk2 knockdown. (G-H) RT-qPCR analysis of stemness-related genes (Nanog, Oct4, Sox2, KLF4, Bmi1) in miR-23b-3p-inhibited LCSCs (G) , and in control, siPyk2, and siPyk2 + miR-23b-3p inhibitor groups (H) . Data are presented as mean ± standard deviation; * P < 0.05, ** P < 0.01, *** P < 0.001; n = 3

Article Snippet: A hydrophobic pen was used to draw a circle around the tissue sections, and endogenous peroxidase activity was blocked, followed by overnight incubation with the primary antibody against Pyk2 (dilution 1:100, 17592-1-AP, Proteintech, Wuhan, China) and Src (dilution 1:50, #2105, Cell signaling technology, Massachusetts, USA) at 4 °C.

Techniques: Expressing, Migration, Transfection, Inhibition, CCK-8 Assay, Knockdown, Quantitative RT-PCR, Control, Standard Deviation

Journal: Cancer Cell International

Article Title: Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression

doi: 10.1186/s12935-025-03841-8

Figure Lengend Snippet: miR-23b promotes HCC progression by regulating Pyk2/Src expression in BALB/c mice. (A-C) HCCLM3 and interfered HCCLM3-LCSCs (control, siPyk2, miR-23b-3p inhibitor, or siPyk2 + miR-23b-3p inhibitor) were subcutaneously injected into BALB/c nude mice ( n = 3 per group). Tumors were excised at the end of the experiment for volume (A) and weight (B) assessment, tumor volume was monitored every 3 days (C) . (D-E) Representative images and quantitative analysis of immunohistochemical staining of Pyk2 and Src in tumor tissues from each group. Scale bar: 50 μm. Data are presented as mean ± standard deviation; * P < 0.05, ** P < 0.01, *** P < 0.001; n = 3

Article Snippet: A hydrophobic pen was used to draw a circle around the tissue sections, and endogenous peroxidase activity was blocked, followed by overnight incubation with the primary antibody against Pyk2 (dilution 1:100, 17592-1-AP, Proteintech, Wuhan, China) and Src (dilution 1:50, #2105, Cell signaling technology, Massachusetts, USA) at 4 °C.

Techniques: Expressing, Control, Injection, Immunohistochemical staining, Staining, Standard Deviation

Journal: PLoS ONE

Article Title: Necl-4/SynCAM-4 Is Expressed in Myelinating Oligodendrocytes but Not Required for Axonal Myelination

doi: 10.1371/journal.pone.0064264

Figure Lengend Snippet: A. Western immuneblotting of brain tissues (from P10 wild-type and Necl-4 homozygous mice) with anti-Necl-1, anti-Necl-2 and anti-Necl-3 antibodies. β-actin was the internal control. B. Statistical analysis on the relative expression level of Necl1, Necl2 and Necl3 with Student's t-test (n = 3. Necl1, p = 0.70. Necl2, p = 0.72. Necl3, p = 0.69). Error bar, standard deviation.

Article Snippet: 30 mg protein from control and mutant tissues was loaded for SDS-PAGE electrophoresis and subsequently detected with anti-Necl-1 (developed in Peking Union Medical University), anti-Necl-2 (Proteintech, Cat# 14335-1-AP), anti-Necl-3 (Abcam Inc, Cat# ab133393) and anti-Necl-4 (UC Davis/NIH NeuroMab Facility, Cat#73-247), and mouse anti-β-actin (Sigma, Cat# A5316) antibodies according to the standard protocol.

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Oxidative medicine and cellular longevity

Article Title: Human Tissue Kallikrein 1 Improves Erectile Dysfunction of Streptozotocin-Induced Diabetic Rats by Inhibition of Excessive Oxidative Stress and Activation of the PI3K/AKT/eNOS Pathway.

doi: 10.1155/2020/6834236

Figure Lengend Snippet: Figure 2: Verification of CSMC and EC from WTR and TGR and the expression of the hKLK1 in EC by immunofluorescence. Panel (a) shows the verification of CSMC through the expression of desmin (magnification: ×200). Panel (b) shows the verification of EC through the expression of CD31, and hKLK1 was only expressed in EC from TGR but not WTR (magnification: ×100). Panel (c) shows the cell purities of EC and CSMC from WTR and TGR through the bar graph. hKLK1: human tissue kallikrein 1; rKLK1: rat tissue kallikrein 1; WTR: wild-type rats; TGR: transgenic rats; EC: endothelial cell; CSMC: cavernous smooth muscle cell; DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: Primary antibodies against hKLK1 (1 : 200; Sigma-Aldrich), CD31 (1 : 100, Boster), and desmin (1 : 200; Abcam) were used in this study.

Techniques: Expressing, Transgenic Assay

Journal: Biomedicines

Article Title: Comparative Proteomic Analysis of tPVAT during Ang II Infusion

doi: 10.3390/biomedicines9121820

Figure Lengend Snippet: List of up-regulated proteins during Ang II infusion.

Article Snippet: The primary antibodies against KDM1A (BM4356), ALCAM (A01788-1), MBNL1 (A02309-1), and TSN (A02777) were purchased from Boster Biological Technology (Wuhan, Hubei, China).

Techniques: Ubiquitin Proteomics, Migration, Membrane

Journal: Biomedicines

Article Title: Comparative Proteomic Analysis of tPVAT during Ang II Infusion

doi: 10.3390/biomedicines9121820

Figure Lengend Snippet: Immunohistochemical verification of DEPs. ( A ) Immunohistochemistry staining of up-regulated proteins, such as lysine-specific histone demethylase 1A (KDM1A), serine/threonine-protein kinase N1 (PKN1), C-terminal-binding protein 1 (CtBP1), transmembrane protein 41B (TMEM41B), myeloblastin (PRTN3), CD166 antigen (ALCAM), and GRB10-interacting GYF protein 2 (GIGYF2). Positive staining was indicated by brown coloration, and nuclei were stained with hematoxylin in blue. ( B ) Immunohistochemistry staining of down-regulated proteins, such as CCN family member 1 (CCN1), muscleblind-like protein 1 (MBNL1), H/ACA ribonucleoprotein complex subunit 2 (NHP2), retinol dehydrogenase 10 (RDH10), T-cell immunoglobulin and mucin domain-containing protein 4 (TIMD4), and Translin (TSN). Positive cells are indicated by brown coloration.

Article Snippet: The primary antibodies against KDM1A (BM4356), ALCAM (A01788-1), MBNL1 (A02309-1), and TSN (A02777) were purchased from Boster Biological Technology (Wuhan, Hubei, China).

Techniques: Immunohistochemical staining, Immunohistochemistry, Staining, Binding Assay

Journal: Biomedicines

Article Title: Comparative Proteomic Analysis of tPVAT during Ang II Infusion

doi: 10.3390/biomedicines9121820

Figure Lengend Snippet: DEPs with high connectivity degree in PPI analysis between saline and Ang II infusion group.

Article Snippet: The primary antibodies against KDM1A (BM4356), ALCAM (A01788-1), MBNL1 (A02309-1), and TSN (A02777) were purchased from Boster Biological Technology (Wuhan, Hubei, China).

Techniques: Saline, Ubiquitin Proteomics

Journal: Cell Death & Disease

Article Title: Deciphering the tumor immune microenvironment of imatinib-resistance in advanced gastrointestinal stromal tumors at single-cell resolution

doi: 10.1038/s41419-024-06571-3

Figure Lengend Snippet: a Cell communication analysis on TIGIT-NECTIN2 and BTLA-TNFRSF14 pair between different cell types in imatinib resistant and sensitive patients respectively. Expression of NECTIN2 ( b ), BTLA ( c ) and TNFRSF14 ( d ) in each cell type in imatinib resistant and sensitive patients respectively. e IHC analysis of NECTIN2, BTLA and TNFRSF14 between imatinib resistant (upper) and sensitive (bottom) patients. f Schematic diagram of the unique tumor-immune microenvironment of imatinib-resistance in advanced GIST.

Article Snippet: Finally, two experienced pathologists independently evaluated staining results for Foxp3 (Servicebio, GB11093), BTLA (Boster, A03149), TNFRSF14 (Affinity, AB2838686) and NECTIN2 (Boster, A08081-2).

Techniques: Expressing