- 701620 - Invitrogen Antibodies TNNT2 antibody

Submitted references Cardiac Tissue-like 3D Microenvironment Enhances Route towards Human Fibroblast Direct Reprogramming into Induced Cardiomyocytes by microRNAs.

Paoletti C, Marcello E, Melis ML, Divieto C, Nurzynska D, Chiono V
Cells 2022 Feb 25;11(5)

MicroRNA-Mediated Direct Reprogramming of Human Adult Fibroblasts Toward Cardiac Phenotype.

Paoletti C, Divieto C, Tarricone G, Di Meglio F, Nurzynska D, Chiono V
Frontiers in bioengineering and biotechnology 2020;8:529

Metabolic Maturation Media Improve Physiological Function of Human iPSC-Derived Cardiomyocytes.

Feyen DAM, McKeithan WL, Bruyneel AAN, Spiering S, Hörmann L, Ulmer B, Zhang H, Briganti F, Schweizer M, Hegyi B, Liao Z, Pölönen RP, Ginsburg KS, Lam CK, Serrano R, Wahlquist C, Kreymerman A, Vu M, Amatya PL, Behrens CS, Ranjbarvaziri S, Maas RGC, Greenhaw M, Bernstein D, Wu JC, Bers DM, Eschenhagen T, Metallo CM, Mercola M
Cell reports 2020 Jul 21;32(3):107925

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of Human cardiomyocytes stained with TNNT2 Recombinant Rabbit Monoclonal Antibody (Product # 701620, 5.0 µg/mL) using a: Alexa Fluor® 488 Goat anti-Rabbit as a secondary antibody (green). b: DAPI stained Primary human cardiomyocytes nuclei (blue). c: Actin stained with Alexa Fluor® 594 Phalloidin (red). d: composite image of cells showing cytoplasmic localization. e: No primary control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 3 Induction of cardiomyocyte markers in miRcombo-transfected AHCFs. (A,B) Gene expression of cardiomyocyte markers TNNT2 and TNNI3 in AHCFs transfected with miRcombo (red) or negmiR (blue) evaluated by ddPCR. Data are representative of three independent experiments, each performed in triplicate. (C) Representative flow plots (left panel) and percentage (right panel) of cTnT positive cells in AHCFs transfected with miRcombo ( n = 3) and negmiR ( n = 3) 15 days after transfection. (D-F) Gene expression of fibroblastic markers VIM , DDR2 , and FSP-1 in AHCFs transfected with miRcombo (red) or negmiR (blue) by ddPCR. Data are representative of three independent experiments, each performed in triplicate. Stated P -value is versus negmiR.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 BM-coated PS plates enhanced AHCF adhesion and miRcombo-mediated direct reprogramming into iCMs. Schematic representations of: ( a ) cell culture conditions on uncoated polystyrene (PS) and BM-coated PS dishes; ( b ) cell transfection (with miRcombo or negmiR) and culture on different conditions for 15 days post-transfection. Image created with Biorender.com under license. ( c ) Cell adhesion on uncoated and BM-coated PS dishes at different time points (0.5, 1, 2 and 4 h): cell adhesion percentage (left panel) and representative images (right panel) of adherent DAPI-stained cells. Experiments were conducted in triplicate, five fields for each sample were taken using a fluorescence microscope. Nuclei were counterstained using DAPI. Scale bar, 100 um. ( d - f ) Expression of TNNT2, ACTC1 and CACNA1C cardiomyocyte genes analyzed by ddPCR in AHCFs transfected with negmiR or miRcombo and cultured on uncoated or BM-coated PS plates for 15 days. Results, expressed as DNA copies/uL, are the average of three independent experiments, each performed in triplicate. ( g ) Representative flow plots of cTnT + cells in AHCFs transfected with negmiR or miRcombo and cultured on BM-coated PS plates ( n = 3) for 15 days. ( h ) Graph showing flow cytometry percentage of cTnT + cells in AHCFs transfected with negmiR or miRcombo and cultured on PS or BM-coated PS plates ( n = 3) for 15 days. All data are expressed as the mean +- SEM. Statistical differences between the groups were determin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Three-dimensional BM hydrogel enhances miRcombo-mediated cell reprogramming as assessed by the analysis of cardiac marker expression by ddPCR analysis and flow cytometry. ( a ) Representative images of different culture conditions: transfected cells are cultured on the top of previously formed hydrogels (2D hydrogel), encapsulated into 3D hydrogel (3D hydrogels) and into 3D hydrogel with BM (3D BM hydrogel) . Image created with Biorender.com under license. ( b ) Representative flow plots of cTnT + cells from AHCFs transfected with negmiR or miRcombo on 2D hydrogel ( n = 3) and cultured for 15 days. ( c ) Graph showing flow cytometry percentage of cTnT + cells from AHCFs transfected with negmiR or miRcombo on PS and 2D hydrogel ( n = 3) and cultured for 15 days. ( d - f ) ddPCR analysis of TNNT2, MYL7 and SCN5A cardiomyocyte gene expression in AHCFs transfected with negmiR or miRcombo and cultured on PS plates, 2D and 3D hydrogels for 15 days. All data are expressed as the mean +- SEM. Statistical differences were determined with two-sided t-tests. ( g - k ) ddPCR analysis of TNNT2, MYL7, SCN5A, ACTC1 and CACNA1C cardiomyocyte genes for miRcombo-transfected cells cultured in PS, BM, 2D hydrogels, 3D hydrogels and 3D BM hydrogels ( n = 3) for 15 days. All data are expressed as the mean +- SEM. Statistical differences are reported between 3D BM hydrogels compared to other culture conditions and were determined with two-sided t-tests. * p < 0.05, ** p < 0.01 and *** p < 0.001.

701620

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