Journal: Biomedical Reports

Article Title: Silencing of lncRNA TFAP2A-AS1 attenuates the development of acute coronary syndrome by inhibiting TFAP2A expression

doi: 10.3892/br.2025.2093

Figure Lengend Snippet: Serum from patients with ACS exhibits elevated expression levels of TFAP2A-AS1 and TFAP2A. The expression of (A) TFAP2A-AS1 and (B) TFAP2A in patients with ACS and healthy individuals was detected by reverse transcription-quantitative PCR. *** P<0.001 vs. Healthy. ACS, acute coronary syndrome; TFAP2A, transcription factor AP-2α.

Article Snippet: Primary antibodies TFAP2A (cat. no. 13019-3-AP), IgG control (cat. no. 30000-0-AP) and GAPDH (cat. no. 60004-1-Ig) were purchased from Proteintech Group, Inc., along with horseradish peroxidase (HRP)-conjugated secondary antibodies (cat. no. SA00001-2).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Biomedical Reports

Article Title: Silencing of lncRNA TFAP2A-AS1 attenuates the development of acute coronary syndrome by inhibiting TFAP2A expression

doi: 10.3892/br.2025.2093

Figure Lengend Snippet: Silencing of TFAP2A-AS1 suppresses TFAP2A expression. (A) The expression of TFAP2A-AS1 in HCAECs after transfection with TFAP2A-AS1 siRNA1/2/3 or NC siRNA was detected by RT-qPCR. *** P<0.001 vs. NC siRNA. (B) The expression of TFAP2A in HCAECs after transfection with TFAP2A siRNA1/2/3 or NC siRNA was detected by RT-qPCR. *** P<0.001 vs. NC siRNA. (C) The expression of TFAP2A-AS1 and TFAP2A in HCAECs after transfection with TFAP2A-AS1 siRNA, TFAP2A siRNA or NC siRNA was detected by RT-qPCR. *** P<0.001 vs. NC siRNA. (D) The protein levels of TFAP2A in HCAECs after transfection with TFAP2A-AS1 siRNA, TFAP2A siRNA or NC siRNA were determined using western blotting. ** P<0.01 and *** P<0.001 vs. NC siRNA. (E) RIP assay demonstrated an enrichment of TFAP2A-AS1. *** P<0.001 vs. anti-IgG. (F) RNA pull-down assay showed that TFAP2A interacting with biotin-labeled TFAP2A-AS1 was higher than that with the antisense of TFAP2A-AS1 group. TFAP2A, transcription factor AP-2α; HCAECs, human coronary artery endothelial cells; siRNA, small interfering RNA; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; RIP, RNA immunoprecipitation; ns, no significance.

Article Snippet: Primary antibodies TFAP2A (cat. no. 13019-3-AP), IgG control (cat. no. 30000-0-AP) and GAPDH (cat. no. 60004-1-Ig) were purchased from Proteintech Group, Inc., along with horseradish peroxidase (HRP)-conjugated secondary antibodies (cat. no. SA00001-2).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Pull Down Assay, Labeling, Small Interfering RNA, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction, RNA Immunoprecipitation

Journal: Biomedical Reports

Article Title: Silencing of lncRNA TFAP2A-AS1 attenuates the development of acute coronary syndrome by inhibiting TFAP2A expression

doi: 10.3892/br.2025.2093

Figure Lengend Snippet: Silencing of TFAP2A-AS1 and TFAP2A suppresses the proliferative, migratory, and invasive capacities while enhancing the apoptotic rate of HCAECs. (A) The apoptosis rate, (B) viability, (C) invasion and (D) migration of HCAECs transfected TFAP2A-AS1 siRNA, TFAP2A siRNA or NC siRNA was assessed by flow cytometric analysis, Counting Kit-8 assay, Transwell invasion assay and wound healing assay, respectively. Scale bar, 100 µm. *** P<0.001 vs. NC siRNA. TFAP2A, transcription factor AP-2α; HCAECs, human coronary artery endothelial cells; siRNA, small interfering RNA; NC, negative control.

Article Snippet: Primary antibodies TFAP2A (cat. no. 13019-3-AP), IgG control (cat. no. 30000-0-AP) and GAPDH (cat. no. 60004-1-Ig) were purchased from Proteintech Group, Inc., along with horseradish peroxidase (HRP)-conjugated secondary antibodies (cat. no. SA00001-2).

Techniques: Migration, Transfection, Transwell Invasion Assay, Wound Healing Assay, Small Interfering RNA, Negative Control

Journal: Biomedical Reports

Article Title: Silencing of lncRNA TFAP2A-AS1 attenuates the development of acute coronary syndrome by inhibiting TFAP2A expression

doi: 10.3892/br.2025.2093

Figure Lengend Snippet: Knockdown of TFAP2A-AS1 and TFAP2A leads to a reduction in serum lipid levels and an improvement in myocardial injury in an ACS mouse model. (A) The expression of TFAP2A-AS1 and TFAP2A in ACS mice after injection of TFAP2A-AS1 shRNA, TFAP2A shRNA or NC shRNA was detected by reverse transcription-quantitative PCR. (B) The protein levels of TFAP2A in ACS mice after injection of TFAP2A-AS1 shRNA, TFAP2A shRNA or NC shRNA were determined using western blotting. (C) The levels of TC, LDL-C and HDL-C in ACS mice after injection of TFAP2A-AS1 shRNA, TFAP2A shRNA or NC shRNA were determined using biochemical tests. (D) Hematoxylin and eosin staining was performed to observe the pathological condition of myocardial tissues in different groups. Scale bar, 50 µm. *** P<0.001 vs. sham; # P<0.05, ## P<0.01, and ### P<0.001 vs. the ACS model + NC shRNA. TFAP2A, transcription factor AP-2α; ACS, acute coronary syndrome; shRNA, short hairpin RNA; ns, no significance.

Article Snippet: Primary antibodies TFAP2A (cat. no. 13019-3-AP), IgG control (cat. no. 30000-0-AP) and GAPDH (cat. no. 60004-1-Ig) were purchased from Proteintech Group, Inc., along with horseradish peroxidase (HRP)-conjugated secondary antibodies (cat. no. SA00001-2).

Techniques: Knockdown, Expressing, Injection, shRNA, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Staining

Journal: Cancer Research

Article Title: PRMT3 Drives IDO1-Dependent Radioresistance and Immunosuppression by Promoting Kynurenine Metabolism in Non–Small Cell Lung Cancer

doi: 10.1158/0008-5472.CAN-24-4162

Figure Lengend Snippet: PRMT3 regulates IDO1 transcription through TFAP2A. A, Co-IP experiments showed that PRMT3 does not directly interact with IDO1. B, Predicted transcription factors of IDO1 intersected with PRMT3-interacting proteins. C and D, PRMT3 interacts with TFAP2A ( C ), and they colocalize ( D ). Scale bar, 20 μm. E, ChIP assays validated the binding of TFAP2A to the IDO1 promoter ( n = 3). F, Luciferase assays indicated that TFAP2A is a transcription factor for IDO1 ( n = 3). G, PRMT3 overexpression promoted the binding of TFAP2A to the IDO1 promoter ( n = 3). H and I, PRMT3 knockdown inhibited TFAP2A binding to the IDO1 promoter, whereas TFAP2A knockdown rescued the PRMT3-mediated upregulation of IDO1 ( n = 3). J, Overexpression of PRMT3 did not increase IDO1 expression in the absence of TFAP2A. K, PRMT3 did not affect TFAP2A mRNA levels ( n = 3). L and M, Overexpression of PRMT3 promoted ADMA generation in TFAP2A ( L ), whereas PRMT3 influenced TFAP2A protein levels ( M ). N, Construction of PRMT3 enzyme inactivation mutant. O, Overexpression of PRMT3 with enzyme inactivation did not increase ADMA production in TFAP2A. P, Observation of the effect of treating cells with PRMT3 inhibitor on ADMA production in TFAP2A. Q, Western blot is used to detect the regulatory effect of enzyme inactivation mutants on TFAP2A protein levels. R, PRMT3 inhibitors can rescue the upregulation of TFAP2A protein levels caused by overexpression of PRMT3. S, There was a positive correlation between PRMT3 and TFAP2A expression in the NSCLC cohort. Data represent the mean ± SD. Differences were tested using unpaired two-sided Student t test ( E and K ) and one-way ANOVA test ( F – I ). S, The correlation was determined using Pearson correlation test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Additionally, GeneChem was responsible for the construction of plasmids aimed at targeting IDO1 (shIDO1 and oeIDO1), TFAP2A (shTFAP2A and oeTFAP2A), and mutants, along with a negative control for comparison.

Techniques: Co-Immunoprecipitation Assay, Binding Assay, Luciferase, Over Expression, Knockdown, Expressing, Mutagenesis, Western Blot

Journal: Cancer Research

Article Title: PRMT3 Drives IDO1-Dependent Radioresistance and Immunosuppression by Promoting Kynurenine Metabolism in Non–Small Cell Lung Cancer

doi: 10.1158/0008-5472.CAN-24-4162

Figure Lengend Snippet: PRMT3-mediated methylation extends TFAP2A protein stability. A and B, PRMT3 overexpression prolonged TFAP2A half-life and promoted nuclear uptake. C, IF revealed that PRMT3 regulates TFAP2A nuclear localization ( n = 3). Scale bar, 20 μm. D, Disuccinimidyl suberate (DSS) assays showed that PRMT3 knockdown inhibited TFAP2A dimerization. E–J, PRMT3 enzyme-inactivated mutants did not affect TFAP2A half-life ( E ), nuclear uptake ( G ), and dimer formation ( I ). The PRMT3 inhibitor SGC707 rescued the effects of PRMT3 overexpression on TFAP2A half-life ( F ), nuclear localization ( H ), and dimer formation ( J ). Data represent the mean ± SD. Differences were tested using unpaired two-sided Student t test ( C ). *, P < 0.05; **, P < 0.01. CHX, cycloheximide.

Article Snippet: Additionally, GeneChem was responsible for the construction of plasmids aimed at targeting IDO1 (shIDO1 and oeIDO1), TFAP2A (shTFAP2A and oeTFAP2A), and mutants, along with a negative control for comparison.

Techniques: Methylation, Over Expression, Knockdown

Journal: Cancer Research

Article Title: PRMT3 Drives IDO1-Dependent Radioresistance and Immunosuppression by Promoting Kynurenine Metabolism in Non–Small Cell Lung Cancer

doi: 10.1158/0008-5472.CAN-24-4162

Figure Lengend Snippet: PRMT3 methylates TFAP2A at R363. A, Molecular docking simulations were conducted to predict the potential methylation sites of TFAP2A. B, In mammals, the amino acid sequence surrounding the R363 site of TFAP2A is highly conserved. A mutant with the substitution R363K was constructed. DBD, DNA-binding domain; TAD, transactivation domain. C, PRMT3 was unable to directly interact with the mutant proteins. D–H, The mutant proteins were incapable of generating ADMA ( D ), and PRMT3 could not regulate their protein levels ( E ), half-life ( F ), dimerization ( G ), or nuclear localization ( H ). CHX, cycloheximide.

Article Snippet: Additionally, GeneChem was responsible for the construction of plasmids aimed at targeting IDO1 (shIDO1 and oeIDO1), TFAP2A (shTFAP2A and oeTFAP2A), and mutants, along with a negative control for comparison.

Techniques: Methylation, Sequencing, Mutagenesis, Construct, Binding Assay