Journal: JCI insight

Article Title: Basigin deficiency prevents anaplerosis and ameliorates insulin resistance and hepatosteatosis.

doi: 10.1172/jci.insight.142464

Figure Lengend Snippet: Figure 3. Bsg deficiency impairs hepatic availability of lactate and pyruvate in vivo. (A) Schematic illustrating impaired gluconeogenesis resulting from decreased substrate import due to Bsg deficiency. OAA, oxaloacetate. (B) Blood glucose levels in Bsg+/+ and Bsg–/– mice under fasting conditions. White columns and circles, Bsg+/+ mice; gray columns and black circles, Bsg–/– mice (n = 10–12/genotype). Scatter plots display the data for individual mice. (C) Dif- ferences in blood lactate values between feeding and fasting states in Bsg+/+ or Bsg–/– female mice (n = 5–6/genotype). (D) Differences in serum pyruvate values between feeding and fasting states in Bsg+/+ or Bsg–/– female mice (n = 5–6/genotype). (E) G6P and F6P with incorporation of 13C3-labeled carbon in isolated Bsg+/+ or Bsg–/– hepatocytes. White columns, Bsg+/+ hepatocytes; gray columns, Bsg–/– hepatocytes. n = 5 for independent experiments. N.D., no detection. (F) Blood glucose excursions and the AUC scores in fasting Bsg+/+ and Bsg–/– female mice during lactate tolerance tests (n = 7–11/genotype). (G) Blood glucose excursions and AUC scores during pyruvate tolerance tests in female mice (n = 8–10/genotype). (H) Endogenous glucose production in isolat- ed hepatocytes of Bsg+/+ and Bsg–/– female mice cultured in medium supplemented with 20 mM sodium lactate and 2 mM sodium pyruvate in the absence or presence of 100 nM AZD3965 (an inhibitor of MCT1 activity). n = 6 for independent experiments. For all relevant panels, data are presented as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, for the comparison of Bsg+/+ and Bsg–/– at the indicated time point (2-tailed unpaired Student’s t test).

Article Snippet: The resulting sections were stained with rabbit monoclonal anti–mouse BSG antibody (Ab) (catalog ab212057; Abcam), rabbit anti–mouse MCT1 Ab (catalog 20139-1-AP; Proteintech), rabbit anti–mouse MCT4 Ab (catalog 22787-1-AP; Proteintech), or FITC-conjugated goat anti–mouse IgG Ab (catalog 115-095- 062; Jackson ImmunoResearch).

Techniques: In Vivo, Labeling, Isolation, Cell Culture, Activity Assay, Comparison

Journal: Pharmaceutical research

Article Title: Participation of Monocarboxylate Transporter 8, But Not P-Glycoprotein, in Carrier-Mediated Cerebral Elimination of Phenytoin across the Blood-Brain Barrier

doi: 10.1007/s11095-021-03003-1

Figure Lengend Snippet: Conditions for RT-PCR

Article Snippet: The blocked membrane was incubated with rabbit-derived anti-MCT8 (20676–1-AP; Proteintech, Rosemont, IL, USA) for approximately 12 h at 4°C or mouse-derived anti-Na + , K + -ATPase α1 antibodies (05–369; Merck) for 2 h at 20°C.

Techniques:

Journal: Pharmaceutical research

Article Title: Participation of Monocarboxylate Transporter 8, But Not P-Glycoprotein, in Carrier-Mediated Cerebral Elimination of Phenytoin across the Blood-Brain Barrier

doi: 10.1007/s11095-021-03003-1

Figure Lengend Snippet: Expression of monocarboxylate transporter 8 (MCT8) mRNA in mouse brain capillaries. RT-PCRwas performed in the absence (−) or presence (+) of reverse transcriptase. Total RNA from the mouse brain was used as a positive control for MCT8 mRNA expression. This image was representative of the images from 3 replicates.

Article Snippet: The blocked membrane was incubated with rabbit-derived anti-MCT8 (20676–1-AP; Proteintech, Rosemont, IL, USA) for approximately 12 h at 4°C or mouse-derived anti-Na + , K + -ATPase α1 antibodies (05–369; Merck) for 2 h at 20°C.

Techniques: Expressing, Positive Control

Journal: Pharmaceutical research

Article Title: Participation of Monocarboxylate Transporter 8, But Not P-Glycoprotein, in Carrier-Mediated Cerebral Elimination of Phenytoin across the Blood-Brain Barrier

doi: 10.1007/s11095-021-03003-1

Figure Lengend Snippet: Protein expression of monocarboxylate transporter 8 (MCT8) in rat tran capillaries. Crude membrane fraction (25 μg) from rat brain capillaries and liver was used for the Western blot analysis of MCT8 proteins. Na+, K+-ATPase α1 was used for the loading control of the crude membrane fraction (21). An arrowhead indicates the expected size of the rat MCT8 proteins (14).

Article Snippet: The blocked membrane was incubated with rabbit-derived anti-MCT8 (20676–1-AP; Proteintech, Rosemont, IL, USA) for approximately 12 h at 4°C or mouse-derived anti-Na + , K + -ATPase α1 antibodies (05–369; Merck) for 2 h at 20°C.

Techniques: Expressing, Western Blot

Journal: Pharmaceutical research

Article Title: Participation of Monocarboxylate Transporter 8, But Not P-Glycoprotein, in Carrier-Mediated Cerebral Elimination of Phenytoin across the Blood-Brain Barrier

doi: 10.1007/s11095-021-03003-1

Figure Lengend Snippet: [l4C]Phenytoin uptake in X. oocytes expressing MCT5 (a), MCT8 (b), or MCT9 (c). [l4C]Phenytoin uptake (0.1 μCi/200μL; 9.09μM) in X. oocytes expressing rat MCT5 (a), MCT8 (b), or MCT9 (c) or in X. oocytes injected with nuclease-free water (a, b, and c) In the absence (−, b) or presence of 400μM unlabeled phenytoin, 1 mM bromosulfophthalein (BSP), or 1 mM desipramine (b). Each open circle represents an individual sample. Each column represents the mean ± S.D. (n = 5–20). **p < 0.01, a significant difference.

Article Snippet: The blocked membrane was incubated with rabbit-derived anti-MCT8 (20676–1-AP; Proteintech, Rosemont, IL, USA) for approximately 12 h at 4°C or mouse-derived anti-Na + , K + -ATPase α1 antibodies (05–369; Merck) for 2 h at 20°C.

Techniques: Expressing, Injection

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Differential regulation of centrosome integrity by DNA damage response proteins.

doi: 10.4161/cc.7.14.6303

Figure Lengend Snippet: Figure 1. MDC1 and BRIT1 localized on centrosome. (A) U2OS cells grown on cover slips were mock transfected, transfected with luciferase siRNA, MDC1 siRNA, and BRIT1 siRNA. At 48 h after transfection, cells were fixed with methanol/acetone (1:1) and costained with antibodies against γ-tubulin and MDC1 or γ-tubulin and BRIT1 followed by Alexa Fluor dye-conjugated secondary antibodies. Nuclei were visualized with 4'6-diamidino-2-phenylin- dole-2HCl (DAPI) staining. Scale bar, 20 μm. (B) Wild-type pEGFP-C2-MDC1 and its various deletion mutants were transfected into the U2OS cell line. At 48 h after transfection, the cells were fixed on cover slips with methanol/ acetone (1:1) at -20°C and stained with mouse anti-γ-tubulin followed by Alexa Fluor dye-conjugated secondary antibodies. Nuclei were visualized with DAPI staining. Scale bar, 20 μm.

Article Snippet: BRIT1 antibody was generated by using a GST-BRIT1 fusion protein synthesized by Proteintech (Chicago, IL).

Techniques: Transfection, Luciferase, Staining

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Differential regulation of centrosome integrity by DNA damage response proteins.

doi: 10.4161/cc.7.14.6303

Figure Lengend Snippet: Figure 2. Deficiency of MDC1 or BRIT1 led to centrosome amplification. (A) U2OS cells grown on cover slips were mock-transfected or transfected with luciferase or MDC1 siRNA. At 48 h after transfection, cells were fixed with methanol/acetone (1:1) and costained with antibodies against γ-tubulin and MDC1 followed by Alexa Fluor dye-conjugated secondary antibodies. Nuclei were visualized with DAPI staining. Scale bar, 20 μm. (B) Bar diagram show- ing that 20% of MDC1-depleted cells contained more than two centrosomes, compared with 3% of luciferase siRNA-treated cells. (C) U2OS cells were treated as described above except that wild-type pEGFP-C2-MDC1 and mutant pEGFP-C2-ΔFHA MDC1 were cotransfected with siRNA to rescue the MDC1- deficient phenotype. Scale bar, 20 μm (D) U2OS cells were grown on cover slips and mock-transfected or transfected with luciferase or MDC1 siRNAs. At 48 h after transfection, cells were treated with 2 mM hydroxyurea for an additional 48 h. Cells were then fixed and costained with antibodies against γ-tubulin and MDC1 followed by Alexa Fluor dye-conjugated secondary antibodies. The percentage of cells containing more than two centrosomes with and without hydroxyurea treatment was determined. (E) U2OS cells grown as described above except that cells were treated with 10 Gy of γ-radiation instead of hydroxyurea. The percentage of cells containing more than two centrosomes after radiation treatment was determined. (F) U2OS cells grown on cover slips were mock-transfected or transfected with luciferase or BRIT1 siRNA. At 48 h after transfection, cells were fixed with methanol/acetone (1:1) and were costained with antibodies against γ-tubulin and BRIT1 followed by Alexa Fluor dye-conjugated secondary antibodies. Nuclei were visualized with DAPI staining. Arrow indicates centrosome amplification with defective cytokinesis.

Article Snippet: BRIT1 antibody was generated by using a GST-BRIT1 fusion protein synthesized by Proteintech (Chicago, IL).

Techniques: Amplification, Transfection, Luciferase, Staining, Mutagenesis

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Differential regulation of centrosome integrity by DNA damage response proteins.

doi: 10.4161/cc.7.14.6303

Figure Lengend Snippet: Figure 3. Depletion of MDC1 or BRIT1 leads to aberrant mitotic spindles and defective cytokinesis. (A) U2OS cells grown on cover slips were mock-trans- fected or transfected with luciferase or MDC1 siRNAs. At 48 h after transfection, cells were fixed and costained with antibodies against α-tubulin and anti MDC1. Cells were washed and subsequently stained with Alexa Fluor dye-conjugated secondary antibodies. Nuclei were visualized with DAPI staining. Scale bar, 20 μm. Mitotic cells with abnormal spindles were captured. (B) Mock-transfected or BRIT1 siRNA-transfected U2OS cells were fixed and costained with antibodies against α-tubulin and BRIT1. Mitotic cells with abnormal spindles were captured. Scale bar, 20 μm. Arrows indicates misaligned spindle that failed to align properly to the equatorial plain, along with a chromosome congression failure at the metaphase. (C) Quantitative analysis of normal and defective mitotic cells observed in luciferase- or BRIT1 siRNA-treated cells. (D) α-tubulin and BubR1 (A, Prometaphase; B, Metaphase; C, Prometaphase; D, Metaphase) or (E) α-tubulin and Mad2 (A, Prometaphase; B, Metaphase; C, Anaphase; D, Metaphase; E, Anaphase) to analyze spindle checkpoint activation. Scale bar, 20 μm. Arrow indicates the persisten accumulation of Mad2 at the metaphase-anaphase transition.

Article Snippet: BRIT1 antibody was generated by using a GST-BRIT1 fusion protein synthesized by Proteintech (Chicago, IL).

Techniques: Transfection, Luciferase, Staining, Activation Assay

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Differential regulation of centrosome integrity by DNA damage response proteins.

doi: 10.4161/cc.7.14.6303

Figure Lengend Snippet: Figure 4. MDC1 and BRIT1 were negative regulators of Aurora A and Plk1. (A) U2OS cells were transfected with luciferase siRNA or MDC1 siRNA (left) or BRIT1 siRNA (right). At 48 h after transfection, cells were harvested and the lysates were subjected to Western blotting using antibodies against (left) MDC1, Plk1 and Aurora A or (right) BRIT1, Plk1 and Aurora A. Actin is for the loading control. (B) U2OS cells were transfected with the siRNA against luciferase, MDC1, or BRIT1. At 48 h after transfection, cells were untreated or treated with cycloheximide for 8 or 12 h. Cells were then harvested, and the lysates were subjected to Western blotting using anti- bodies against Plk1, Aurora A or actin. (C) U2OS cells were transfected with siRNA against luciferase or MDC1 along with cotransfection of empty expression vector or the vectors express- ing wild-type pEGFP-C2-MDC1 or pEGFP-C2-ΔFHA-MDC1 mutant. At 48 h after the transfection, cells were harvested, and the lysates were subjected to Western blotting using antibodies against MDC1, Plk1, Aurora A. Actin is used for the loading control.

Article Snippet: BRIT1 antibody was generated by using a GST-BRIT1 fusion protein synthesized by Proteintech (Chicago, IL).

Techniques: Transfection, Luciferase, Western Blot, Control, Cotransfection, Expressing, Plasmid Preparation, Mutagenesis

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Differential regulation of centrosome integrity by DNA damage response proteins.

doi: 10.4161/cc.7.14.6303

Figure Lengend Snippet: Figure 5. Expression levels of MDC1 and BRIT1 correlated with centrosome amplification and disease progression in breast cancer specimens. Paraffin-embedded adjacent normal and breast cancer specimens were analyzed by immunohistochemical staining. (A) MDC1 and (B) BRIT1 protein expression was correlated with the intensity of brown color in normal and breast cancer specimens. Scale bar, 50 mm. Frozen adjacent normal and breast cancer specimens were immu- nostained using antibodies against (C) γ-tubulin and MDC1 or (D) Plk1 and BRIT1. Cells were washed and subsequently stained with Alexa Fluor dye-conjugated secondary antibodies. Cells containing more than two centrosomes and mitotic cells with abnormal spindles were visualized. Scale bar, 20 μm. Arrow indicates the overexpression of Plk1 in breast cancer as compared to normal breast specimen. (E) MDC1 mRNA levels from breast cancer were analyzed for correlation with time of freedom from metastasis in a database maintained by Stanford University. Patients whose cancer samples expressed normal to high MDC1 levels were associated with a longer time to develop metastasis and an overall reduction in the incidence of metastasis.

Article Snippet: BRIT1 antibody was generated by using a GST-BRIT1 fusion protein synthesized by Proteintech (Chicago, IL).

Techniques: Expressing, Amplification, Biomarker Discovery, Immunohistochemical staining, Staining, Over Expression

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Differential regulation of centrosome integrity by DNA damage response proteins.

doi: 10.4161/cc.7.14.6303

Figure Lengend Snippet: Figure 6. Model illustrating the role of MDC1 and BRIT1 in maintaining centrosome integrity and cytokinesis. MDC1 and BRIT1 are both required for maintaining numerical centrosome homeostasis, mitotic spindle assembly, and cytokinesis by negatively regulating Aurora A and Plk1. Deficiency of MDC1 led to centrosome amplification followed by multipolar mitosis and chromosome misseg- regation resulting in aneuploidy. BRIT1 depletion led to misaligned spindles followed by spindle checkpoint activation. Because BRIT1 depletion also led to increased Aurora A and Plk1, the spindle checkpoint was eventually overcome and, in turn, triggered defective cytokinesis, leading to the generation of one polyploid cell.

Article Snippet: BRIT1 antibody was generated by using a GST-BRIT1 fusion protein synthesized by Proteintech (Chicago, IL).

Techniques: Amplification, Activation Assay

Journal: Cell Death Discovery

Article Title: MCT4-driven CAF-mediated metabolic reprogramming in breast cancer microenvironment is a vulnerability targetable by miR-425-5p

doi: 10.1038/s41420-024-01910-x

Figure Lengend Snippet: Representative Western blot analysis of MCT4 performed on plasmamembrane fractions of Cancer-Associated Fibroblasts (CAFs) ( n = 4) and Normal Fibroblasts (NFs) ( n = 3). Na + /K+ ATPase was used for data normalization of the fractionation in the sample preparation. Densitometry data are expressed after normalization on Na + /K+ ATPase band intensity A . A representative confocal microscopy image of MCT4 expression in CAFs ( n = 3) (DAPI blue, MCT4 red) B . Three different subpopulations of two primary fibroblasts were collected at different growing distances from the tumor (intratumoral, IT; peritumoral, PT; normal, NF) and MCT4 gene expression was evaluated by quantitative real-time PCR. Values are expressed as a mean of two biological replicates of technical duplicates C . Culture media from NFs ( n = 2) and CAFs ( n = 2) were collected and assessed by a colorimetric L-lactate assay. Values are expressed as mean of two technical replicates D . Graph shows mean ± SD. * P values < 0.05; *** p < 0.001. Bar = 100 μm.

Article Snippet: Plasmid transfection in MS-5 (2500 ng of SLC16A3 tGFP-tagged supplied by Origene CAT#: RG211987) was carried out using Lipofectamine 3000 reagent (Invitrogen).

Techniques: Western Blot, Fractionation, Sample Prep, Confocal Microscopy, Expressing, Real-time Polymerase Chain Reaction, Lactate Assay

Journal: Cell Death Discovery

Article Title: MCT4-driven CAF-mediated metabolic reprogramming in breast cancer microenvironment is a vulnerability targetable by miR-425-5p

doi: 10.1038/s41420-024-01910-x

Figure Lengend Snippet: Bioinformatics predictions (TargetScan and miRANDA) indicated miR-425-5p as a putative miRNA targeting MCT4 mRNA A . Representative Real Time PCR for miR-425-5p upon miR-425-5p and anti miR-425-5p transfection in Cancer-Associated Fibroblasts (CAF n = 1) and Normal Fibroblasts (NF n = 1) respectively. Data are expressed as mean of two technical replicates B . Representative Western Blot analysis of MCT4 upon miR-425-5p and anti-miR-425 transfection in CAFs ( n = 4) and NFs ( n = 2) respectively C . Dual luciferase assay of the predicted binding site for miR-425-5p on the 3’UTR region of SLC16A3. Data are expressed as mean of two technical replicates D . Quantitative Real Time PCR for miR-425-5p performed on CAFs ( n = 3) and NFs ( n = 2), to evaluate miR-425-5p levels. Values are expressed as mean of two technical replicates E . Graph shows mean ± SD over control. * P values < 0.05.

Article Snippet: Plasmid transfection in MS-5 (2500 ng of SLC16A3 tGFP-tagged supplied by Origene CAT#: RG211987) was carried out using Lipofectamine 3000 reagent (Invitrogen).

Techniques: Real-time Polymerase Chain Reaction, Transfection, Western Blot, Luciferase, Binding Assay, Control

Journal: Cell Death Discovery

Article Title: MCT4-driven CAF-mediated metabolic reprogramming in breast cancer microenvironment is a vulnerability targetable by miR-425-5p

doi: 10.1038/s41420-024-01910-x

Figure Lengend Snippet: Metabolic changes in MCT4-transfected MS-5 cells with or without miR-425-5p overexpression, as assessed by glycolytic proton efflux rate (glycoPER) kinetics A , proton efflux rate (PER) B , extracellular acidification rate (ECAR) C , oxygen consumption rate OCR D , and ATP production E . Data are presented as a mean of biological duplicates of eight technical replicates ± SD over control.

Article Snippet: Plasmid transfection in MS-5 (2500 ng of SLC16A3 tGFP-tagged supplied by Origene CAT#: RG211987) was carried out using Lipofectamine 3000 reagent (Invitrogen).

Techniques: Transfection, Over Expression, Control

Journal: Cell Death Discovery

Article Title: MCT4-driven CAF-mediated metabolic reprogramming in breast cancer microenvironment is a vulnerability targetable by miR-425-5p

doi: 10.1038/s41420-024-01910-x

Figure Lengend Snippet: MiR-425-5p alters the metabolism of breast Cancer-associated Fibroblasts (CAFs) by downregulating MCT4 and indirectly reducing GLUT1, leading to decreased lactate extrusion. The interaction between miR-425-5p reprogrammed CAFs and breast cancer epithelial cells has a significant impact on breast cancer cell viability, proliferation, and migration. Angiogenesis is also influenced. This graphical abstract was drawn by using and adapting pictures from Servier Medical Art (Smart.Servier.com), provided by Servier, licensed under a Creative Commons Attribution 3.0 unported license ( http://creativecommons.org/licenses/by/3.0/ ).

Article Snippet: Plasmid transfection in MS-5 (2500 ng of SLC16A3 tGFP-tagged supplied by Origene CAT#: RG211987) was carried out using Lipofectamine 3000 reagent (Invitrogen).

Techniques: Migration

Journal: Journal of Inflammation Research

Article Title: Exploring Long Non-Coding RNAs Associated with IP3/DAG Signaling Pathway as Potential Biomarkers Involved in Mast Cell Degranulation in Chronic Spontaneous Urticaria with 2-Year Follow-Up

doi: 10.2147/JIR.S343826

Figure Lengend Snippet: Serum concentrations of hs-CRP, LTB4, PGD2, MCT, D-dimer and HIS in the CSU (n = 56) and healthy control (n = 13) groups. Differences between groups were assessed using Mann–Whitney U -tests; experiments were repeated three times.

Article Snippet: The levels of HIS (ab213975, Abcam, Cambridge, UK), MCT (CSB-E09012h, CUSABIO, Wuhan, China), LTB4 (CSB-E08033h), PGD2 (CSB-E13898h), hs-CRP (CSB-E08617h), and D-dimer (CSB-E05175h) were assessed in peripheral blood serum using human ELISA kits according to the manufacturer’s instructions.

Techniques: Control, MANN-WHITNEY

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – Sequência de iniciadores para SLC26A4-AS1 e GAPDH

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques:

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 é regulado para cima na hipertrofia cardíaca induzida por AngII. (A-C) Em NMVCs tratados com 150 nM AngII por 1, 6, 12 e 24 h, a expressão de biomarcadores hipertróficos (ANP, BNP e β-MHC) foi avaliada usando RT-qPCR, e o nível de proteína desses biomarcadores foi avaliada por western blot, seguida de quantificação por western blot. *P<0,05: AngII (1 h) vs AngII (0 h). *P<0,05: AngII (6 h) vs AngII (0 h). **P<0,01: AngII (12 h) vs AngII (0 h). **P<0,01: AngII (24 h) vs AngII (0 h). O número de tamanho da amostra (n) = 3. (D) A área de superfície celular em NMVCs tratadas com 150 nM AngII por 1, 6, 12 e 24 h foi avaliada usando coloração IF. Barra de escala: 10 μm. *P<0,05: AngII (1 h) vs AngII (0 h). *P<0,05: AngII (6 h) vs AngII (0 h). **P<0,01: AngII (12 h) vs AngII (0 h). **P<0,01: AngII (24 h) vs AngII (0 h). n = 3. (E) A expressão de SLC26A4-AS1 em NMVCs tratados com 150 nM AngII por 1, 6, 12 e 24 h foi detectada usando RT-qPCR. *P<0,05: AngII (1 h) vs AngII (0 h). *P<0,05: AngII (6 h) vs AngII (0 h). **P<0,01: AngII (12 h) vs AngII (0 h). **P<0,01: AngII (24 h) vs AngII (0 h). n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Quantitative RT-PCR, Western Blot

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – O silenciamento de SLC26A4-AS1 suprime a hipertrofia cardíaca induzida por AngII. (A) NMVCs foram tratados com sh-SLC26A4-AS1#1/2 ou sh-NC após o tratamento de AngII por 24 h. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1/2 vs AngII+sh-NC. n = 3. (B) A área da superfície celular foi detectada em NMVCs tratados com AngII após o silêncio de SLC26A4-AS1 usando coloração IF. Barra de escala: 10 μm. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1/2 vs AngII+sh-NC. n = 3. (C-H) Após o silenciamento de SLC26A4-AS1, a expressão de biomarcadores hipertróficos em NMVCs tratados com AngII foi medida usando RT-qPCR, e o nível desses biomarcadores foi avaliado usando western blot, seguido pela quantificação de western blot. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1/2 vs AngII+sh-NC. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Quantitative RT-PCR, Western Blot

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 funciona como um ceRNA para regular SLC26A4 na hipertrofia cardíaca induzida por AngII. (A-B) Fracionamento subcelular e ensaios de FISH foram realizados para determinar a localização de SLC26A4-AS1 em NMVCs induzidos por AngII. n = 3. (C) Ensaios RIP detectaram o enriquecimento de SLC26A4-AS1 e SLC26A4. **P<0,01: Ago2 vs IgG. n = 3. (D) A expressão de SLC26A4 em NMVCs tratados com 150 nM AngII por 1, 6, 12 e 24 h foi avaliada usando RT-qPCR. *P<0,05: AngII (1 h) vs AngII (0 h). *P<0,05: AngII (6 h) vs AngII (0 h). **P<0,01: AngII (12 h) vs AngII (0 h). **P<0,01: AngII (24 h) vs AngII (0 h). n = 3. (E-F) O nível de expressão e o nível de proteína de SLC26A4 em NMVCs tratados com AngII foram avaliados usando RT-qPCR e western blot, respectivamente, após regulação negativa de SLC26A4-AS1, seguido pela quantificação de western blot. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1/2 vs AngII+sh-NC. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Quantitative RT-PCR, Western Blot

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 promove hipertrofia cardíaca induzida por AngII via regulação para cima de SLC26A4. (A) NMVCs introduzidos com pcDNA3.1-SLC26A4, ou pcDNA3.1 foram tratados com AngII por 24 h. **P<0,01: pcDNA3.1-SLC26A4 vs pcDNA3.1. Experimentos de resgate foram conduzidos em NMVCs com a transfecção de sh-NC, sh-SLC26A4-AS1#1, sh-SLC26A4-AS1#1+pcDNA3.1 e sh-SLC26A4-AS1#1+pcDNA3.1-SLC26A4 sob tratamento com AngII . n = 3. (B) A área da superfície celular foi avaliada em NMVCs tratados com AngII usando coloração IF. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1 vs AngII+sh-NC. **P<0,01: AngII+sh-SLC26A4-AS1#1+pcDNA3.1-SLC26A4 vs AngII+sh-SLC26A4-AS1#1+pcDNA3.1. n = 3. (C-E) O nível de expressão e o nível de proteína de biomarcadores hipertróficos em NMVCs tratados com AngII foram medidos por meio de RT-qPCR e western blot, respectivamente, seguidos pela quantificação de western blot. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1 vs AngII+sh-NC. **P<0,01: AngII+sh-SLC26A4-AS1#1+pcDNA3.1-SLC26A4 vs AngII+sh-SLC26A4-AS1#1+pcDNA3.1. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: IF-P, Quantitative RT-PCR, Western Blot

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 regula para cima a expressão de SLC26A4 por meio de absorção de miR-301a-3p ou miR-301b-3p. (A) Os miRNAs candidatos combinados com SLC26A4-AS1 e SLC26A4 foram obtidos no site da starBase (http://starbase.sysu.edu.cn). (B) O enriquecimento relativo de miRNAs candidatos em grupos bio-SLC26A4-AS1 foi analisado por meio do ensaio de RNA pull-down. **P<0,01: Bio-SLC26A4-AS1 vs Bio-NC. n = 3. (C) O ensaio RIP detectou o enriquecimento relativo de SLC26A4-AS1, miR-301a-3p, miR-301b-3p e SLC26A4. **P<0,01: Ago2 vs IgG. n = 3. (D-E) A expressão de miR-301a-3p ou miR-301b-3p em NMVCs tratados com 150 nM AngII por 1, 6, 12 e 24 h foi detectada usando RT-qPCR. *P<0,05: AngII (1h)/AngII (6h) vs AngII (0 h). **P<0,01: AngII (12h)/AngII (24h) vs AngII (0 h). n = 3. (F) Ensaios de luciferase repórter foram implementados para detectar a atividade de luciferase de SLC26A4-AS1 do tipo selvagem (WT) ou do tipo mutante (MUT). **P<0,01: mímicos miR-301a-3p/mímicos miR-301b-3p vs mímicos NC. n = 3. (G) Ensaios de repórter de luciferase foram realizados para detectar a atividade de luciferase de SLC26A4 3’UTR WT ou MUT. **P<0,01: mímicos miR-301a-3p/mímicos miR-301b-3p vs miméticos NC. n = 3. (H) Os efeitos da superexpressão de miR-301a-3p ou miR-301b-3p na expressão de SLC26A4 foram avaliados via RT-qPCR. **P<0,01: AngII vs Controle. **P<0,01: miméticos AngII+miR-301a-3p/miméticos miR-301b-3p vs miméticos AngII+NC. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Quantitative RT-PCR, Luciferase

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 afeta a hipertrofia cardíaca induzida por AngII via absorção de miR-301a-3p ou miR-301b-3p. (A-B) NMVCs tratados com inibidor de miR-301a-3p/inibidor de miR-301b-3p ou inibidor de NC foram tratados com AngII por 24 h. **P<0,01: inibidor de miR-301a-3p/inibidor de miR-301b-3p vs inibidor de NC. Experimentos de resgate foram conduzidos em NMVCs com a transfecção de sh-NC, sh-SLC26A4-AS1#1, sh-SLC26A4-AS1#1+inibidor de NC, sh-SLC26A4-AS1#1+miR-301a-3p inibidor e sh -SLC26A4-AS1#1+miR-301b-3p inibidor sob tratamento com AngII. n = 3. (C) A área da superfície celular foi medida em NMVCs tratados com AngII usando coloração IF. Barra de escala: 10 μm. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1 vs AngII+sh-NC. *P<0,05: AngII+sh-SLC26A4-AS1#1+inibidor de miR-301a-3p/inibidor de miR-301b-3p vs AngII+sh-SLC26A4-AS1#1+inibidor de NC. n = 3. (D-F) RT-qPCR e western blot avaliaram o nível de expressão e o nível de proteína de biomarcadores hipertróficos em NMVCs tratados com AngII, respectivamente, seguido pela quantificação de western blot. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1 vs AngII+sh-NC. *P<0,05: AngII+sh-SLC26A4-AS1#1+inibidor de miR-301a-3p/inibidor de miR-301b-3p vs AngII+sh-SLC26A4-AS1#1+inibidor de NC. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Quantitative RT-PCR, Western Blot

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – Primer sequence for SLC26A4-AS1 and GAPDH

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Sequencing

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 is upregulated in AngII-induced cardiac hypertrophy. (A-C) In NMVCs treated with 150 nM AngII for 1, 6, 12, and 24 h, the expression of hypertrophic biomarkers (ANP, BNP, and β-MHC) was assessed using RT-qPCR, and the protein level of these biomarkers was evaluated through western blot, followed by western blot quantification. *P<0.05: AngII (1 h) vs AngII (0 h). *P<0.05: AngII (6 h) vs AngII (0 h). **P<0.01: AngII (12 h) vs AngII (0 h). **P<0.01: AngII (24 h) vs AngII (0 h). The number of sample size (n) = 3. (D) The cell surface area in NMVCs treated with 150 nM AngII for 1, 6, 12, and 24 h was assessed using IF staining. Scale bar: 10 μm. *P<0.05: AngII (1 h) vs AngII (0 h). *P<0.05: AngII (6 h) vs AngII (0 h). **P<0.01: AngII (12 h) vs AngII (0 h). **P<0.01: AngII (24 h) vs AngII (0 h). n = 3. (E) SLC26A4-AS1 expression in NMVCs treated with 150 nM AngII for 1, 6, 12, and 24 h was detected using RT-qPCR. *P<0.05: AngII (1 h) vs AngII (0 h). *P<0.05: AngII (6 h) vs AngII (0 h). **P<0.01: AngII (12 h) vs AngII (0 h). **P<0.01: AngII (24 h) vs AngII (0 h). n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Staining

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 silence suppresses AngII-induced cardiac hypertrophy. (A) NMVCs were treated with sh-SLC26A4-AS1#1/2 or sh-NC after the treatment of AngII for 24 h. **P<0.01: AngII vs Control. **P<0.01: AngII+sh-SLC26A4-AS1#1/2 vs AngII+sh-NC. n = 3. (B) Cell surface area was detected in AngII-treated NMVCs upon SLC26A4-AS1 silence using IF staining. Scale bar: 10 μm. **P<0.01: AngII vs Control. **P<0.01: AngII+sh-SLC26A4-AS1#1/2 vs AngII+sh-NC. n = 3. (C-H) After SLC26A4-AS1 silencing, the expression of hypertrophic biomarkers in AngII-treated NMVCs was measured using RT-qPCR, and the level of these biomarkers was assessed using western blot, followed by the quantification of western blot. **P<0.01: AngII vs Control. **P<0.01: AngII+sh-SLC26A4-AS1#1/2 vs AngII+sh-NC. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Control, Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 functions as a ceRNA to regulate SLC26A4 in cardiac hypertrophy induced by AngII. (A-B) Subcellular fractionation and FISH assays were performed to determine the location of SLC26A4-AS1 in AngII-induced NMVCs. n = 3. (C) RIP assays detected the enrichment of SLC26A4-AS1 and SLC26A4. **P<0.01: Ago2 vs IgG. n = 3. (D) The expression of SLC26A4 in NMVCs treated with 150 nM AngII for 1, 6, 12, and 24 h was assessed using RT-qPCR. *P<0.05: AngII (1 h) vs AngII (0 h). *P<0.05: AngII (6 h) vs AngII (0 h). **P<0.01: AngII (12 h) vs AngII (0 h). **P<0.01: AngII (24 h) vs AngII (0 h). n = 3. (E-F) Expression level and protein level of SLC26A4 in AngII-treated NMVCs were assessed using RT-qPCR and western blot, respectively, after SLC26A4-AS1 down-regulation, followed by the quantification of western blot. **P<0.01: AngII vs Control. **P<0.01: AngII+sh-SLC26A4-AS1#1/2 vs AngII+sh-NC. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Fractionation, Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 promove hipertrofia cardíaca induzida por AngII via regulação para cima de SLC26A4. (A) NMVCs introduzidos com pcDNA3.1-SLC26A4, ou pcDNA3.1 foram tratados com AngII por 24 h. **P<0,01: pcDNA3.1-SLC26A4 vs pcDNA3.1. Experimentos de resgate foram conduzidos em NMVCs com a transfecção de sh-NC, sh-SLC26A4-AS1#1, sh-SLC26A4-AS1#1+pcDNA3.1 e sh-SLC26A4-AS1#1+pcDNA3.1-SLC26A4 sob tratamento com AngII . n = 3. (B) A área da superfície celular foi avaliada em NMVCs tratados com AngII usando coloração IF. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1 vs AngII+sh-NC. **P<0,01: AngII+sh-SLC26A4-AS1#1+pcDNA3.1-SLC26A4 vs AngII+sh-SLC26A4-AS1#1+pcDNA3.1. n = 3. (C-E) O nível de expressão e o nível de proteína de biomarcadores hipertróficos em NMVCs tratados com AngII foram medidos por meio de RT-qPCR e western blot, respectivamente, seguidos pela quantificação de western blot. **P<0,01: AngII vs Controle. **P<0,01: AngII+sh-SLC26A4-AS1#1 vs AngII+sh-NC. **P<0,01: AngII+sh-SLC26A4-AS1#1+pcDNA3.1-SLC26A4 vs AngII+sh-SLC26A4-AS1#1+pcDNA3.1. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: IF-P, Quantitative RT-PCR, Western Blot

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 positively regulates SLC26A4 expression via sponging miR-301a-3p or miR-301b-3p. (A) Candidate miRNAs combined with SLC26A4-AS1 and SLC26A4 were obtained from the starBase website (http://starbase.sysu.edu.cn). (B) The relative enrichment of candidate miRNAs in bio-SLC26A4-AS1 groups was analyzed via RNApull-down assay. **P<0.01: Bio-SLC26A4-AS1 vs Bio-NC. n = 3. (C) The RIP assay detected the relative enrichment of SLC26A4-AS1, miR-301a-3p, miR-301b-3p, and SLC26A4. **P<0.01: Ago2 vs IgG. n = 3. (D-E) Expression of miR-301a-3p or miR-301b-3p in NMVCs treated with 150 nM AngII for 1, 6, 12, and 24 h was detected using RT-qPCR. *P<0.05: AngII (1h)/AngII (6h) vs AngII (0 h). **P<0.01: AngII (12h)/AngII (24h) vs AngII (0 h). n = 3. (F) Luciferase reporter assays were implemented to detect the luciferase activity of SLC26A4-AS1 wild-type (WT) or mutant-type (MUT). **P<0.01: miR-301a-3p mimics/miR-301b-3p mimics vs NC mimics. n = 3. (G) Luciferase reporter assays were performed to detect the luciferase activity of SLC26A4 3’UTR WT or MUT. **P<0.01: miR-301a-3p mimics/miR-301b-3p mimics vs NC mimics. n = 3. (H) The effects of miR-301a-3p or miR-301b-3p overexpression on SLC26A4 expression were assessed via RT-qPCR. **P<0.01: AngII vs Control. **P<0.01: AngII+miR-301a-3p mimics/miR-301b-3p mimics vs AngII+NC mimics. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Expressing, Quantitative RT-PCR, Luciferase, Activity Assay, Mutagenesis, Over Expression, Control

Journal: Arquivos Brasileiros de Cardiologia

Article Title: SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4

doi: 10.36660/abc.20210933

Figure Lengend Snippet: – SLC26A4-AS1 affects AngII-induced cardiac hypertrophy via sponging miR-301a-3p or miR-301b-3p. (A-B) NMVCs introduced with miR-301a-3p inhibitor/miR-301b-3p inhibitor, or NC inhibitor were treated with AngII for 24 h. **P<0.01: miR-301a-3p inhibitor/miR-301b-3p inhibitor vs NC inhibitor. Rescue experiments were conducted in NMVCs with the transfection of sh-NC, sh-SLC26A4-AS1#1, sh-SLC26A4-AS1#1+NC inhibitor, sh-SLC26A4-AS1#1+miR-301a-3p inhibitor, and sh-SLC26A4-AS1#1+miR-301b-3p inhibitor under AngII treatment. n = 3. (C) Cell surface area was measured in AngII-treated NMVCs using IF staining. Scale bar: 10 μm. **P<0.01: AngII vs Control. **P<0.01: AngII+sh-SLC26A4-AS1#1 vs AngII+sh-NC. *P<0.05: AngII+sh-SLC26A4-AS1#1+miR-301a-3p inhibitor/miR-301b-3p inhibitor vs AngII+sh-SLC26A4-AS1#1+NC inhibitor. n = 3. (D-F) RT-qPCR and western blot evaluated the expression level and protein level of hypertrophic biomarkers in AngII-treated NMVCs, respectively, followed by the quantification of western blot. **P<0.01: AngII vs Control. **P<0.01: AngII+sh-SLC26A4-AS1#1 vs AngII+sh-NC. *P<0.05: AngII+sh-SLC26A4-AS1#1+miR-301a-3p inhibitor/miR-301b-3p inhibitor vs AngII+sh-SLC26A4-AS1#1+NC inhibitor. n = 3.

Article Snippet: Subsequently, the membrane was cultured with primary antibodies against ANP (1μg/ml, ab237632, Abcam, Cambridge, MA, USA), BNP (1μg/ml, ab92500, Abcam), β-MHC (1/2500, ab55152, Abcam), SLC26A4 (0.25 μg/μl, 20889-1-AP, Proteintech, Chicago, USA) and GAPDH (1/2500, ab8245, Abcam) at 4°C overnight.

Techniques: Transfection, Staining, Control, Quantitative RT-PCR, Western Blot, Expressing

Journal: Cell reports

Article Title: The intrinsically disordered region of eIF5B stimulates IRES usage and nucleates biological granule formation

doi: 10.1016/j.celrep.2023.113283

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: RC200224 , Origene , MCTS1 (Myc-DDK-tagged)-Human malignant T cell amplified sequence 1 (MCTS1) expression vector.

Techniques: Recombinant, Modification, Transfection, Virus, Lysis, Protein Purification, Plasmid Preparation, Expressing, Luciferase, Amplification, Sequencing, Software