rabbit polyclonal anti terf2  (Novus Biologicals)

Journal: Experimental & molecular medicine

Article Title: SUMOylation of TP53INP1 is involved in miR-30a-5p-regulated heart senescence.

doi: 10.1038/s12276-024-01347-3

Figure Lengend Snippet: Fig. 1 miR-30a-5p is downregulated in aging hearts and NRCMs. a qRT‒PCR analysis of miR-30a-5p expression in the heart tissues of 3- and 18-month-old WT mice (n = 6 in each group). Representative RNA-FISH images (b) and analysis (c) of miR-30a-5p levels in heart tissues of 3- and 18-month-old WT mice (n = 4 in each group). d qRT‒PCR analysis of miR-30a-5p levels in the hearts of vehicle- or D-gal-treated WT mice (n = 5 in each group). Representative RNA-FISH images (e) and analysis (f) of miR-30a-5p levels in the heart tissues of vehicle- or D-gal-induced mice (n = 3 in each group). Representative western blotting images (g) and analysis (h) of p53 and p21 expression in the heart tissues of 3- and 18-month-old WT mice (n = 4 in each group). Representative western blotting images (i) and analysis (j) of p53 and p21 protein levels in the heart tissues of the vehicle- or D-gal-treated mice (n = 4 in each group). k qRT‒PCR analysis of miR-30a-5p expression in the blood of 3- or 18-month-old mice (n = 4 in both groups). l qRT‒PCR analysis of miR-30a-5p expression in NRCMs and NRCFs (n = 6 in each group). m qRT‒ PCR analysis of miR-30a-5p expression in 5- and 10-day-treated NRCMs (n = 9 in each group). n qRT‒PCR analysis of miR-30a-5p expression in vehicle- and D-gal-induced NRCMs (n = 10 in each group). The data are presented as the means ± standard errors. Statistical significance was assessed using t tests. WT wild-type, qRT‒PCR quantitative real-time polymerase chain reaction, RNA‒FISH RNA‒fluorescence in situ hybridization, veh vehicle, D‒gal D‒galactose, NRCMs neonatal rat cardiomyocytes, NRCFs neonatal rat cardiac fibroblasts.

Article Snippet: The primary antibodies used for western blotting were mouse monoclonal anti-p53 (Cat# Ab26, Abcam, Cambridge, UK), rabbit polyclonal anti-p21 (Cat# 8248-1-AP) and mouse monoclonal anti-TERF2 (Cat# 66893- 1-Ig, both ProteinTech, Wuhan, China), rabbit polyclonal anti-TP53INP1 (Cat# OM204116, OmniMabs, Alhambra, CA, USA), mouse monoclonal antiSUMO1 (Cat# sc-5308, Santa Cruz, Dallas, Texas, USA), rabbit polyclonal anti-TERT (Cat# DF7129), rabbit polyclonal anti-DDIT4 (Cat# 10638-1-AP, Proteintech, Wuhan, China), and rabbit polyclonal anti-SENP1 (Cat# AF0275, both Affinity Bioscience, Nanjing, China).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, In Situ Hybridization

Journal: Experimental & molecular medicine

Article Title: SUMOylation of TP53INP1 is involved in miR-30a-5p-regulated heart senescence.

doi: 10.1038/s12276-024-01347-3

Figure Lengend Snippet: Fig. 2 miR-30a-5p KO aggravates the aging-induced decrease in cardiac function and promotes senescence. Representative ECG images (a) and analysis (b) of LVEF and LVFS in 2- and 18-month-old WT and KO mice (n = 7 in each group). Representative images of immunofluorescence staining (c) and analysis (d) of γH2AX in heart tissues of 2- and 18-month-old WT and KO mice (scale bars=200 μm; n = 3 in each group). e qRT‒PCR analysis of telomere length in heart tissues of 2- and 18-month-old WT and KO mice (n = 6 in each group). Representative western blotting images (f) and analysis (g) of the expression of p21 (n = 5 in each group) and p53 (n = 4 in each group) in the heart tissues of 2- and 18-month-old WT and KO mice. qRT‒PCR analysis of myocardial IL-1β (h, n = 5 in each group) and IL-6 (i, n = 4 in each group) levels. Representative Picrosirius Red (scale bars = 50 μm) and wheat germ agglutinin (WGA, scale bars = 100 μm) staining images (j) and analysis (k) of the hearts of 2- and 18-month-old WT and KO mice (n = 5 in each group). The data are presented as the means ± standard errors. Statistical significance was assessed using one-way ANOVA. ECG echocardiography, WT wild-type, KO knockout, LVEF left ventricular ejection fraction, LVFS left ventricular fractional shortening, qRT‒PCR quantitative real-time polymerase chain reaction, γH2AX H2A histone family member X, WGA wheat germ agglutinin.

Article Snippet: The primary antibodies used for western blotting were mouse monoclonal anti-p53 (Cat# Ab26, Abcam, Cambridge, UK), rabbit polyclonal anti-p21 (Cat# 8248-1-AP) and mouse monoclonal anti-TERF2 (Cat# 66893- 1-Ig, both ProteinTech, Wuhan, China), rabbit polyclonal anti-TP53INP1 (Cat# OM204116, OmniMabs, Alhambra, CA, USA), mouse monoclonal antiSUMO1 (Cat# sc-5308, Santa Cruz, Dallas, Texas, USA), rabbit polyclonal anti-TERT (Cat# DF7129), rabbit polyclonal anti-DDIT4 (Cat# 10638-1-AP, Proteintech, Wuhan, China), and rabbit polyclonal anti-SENP1 (Cat# AF0275, both Affinity Bioscience, Nanjing, China).

Techniques: Staining, Western Blot, Expressing, Knock-Out, Real-time Polymerase Chain Reaction

Journal: Experimental & molecular medicine

Article Title: SUMOylation of TP53INP1 is involved in miR-30a-5p-regulated heart senescence.

doi: 10.1038/s12276-024-01347-3

Figure Lengend Snippet: Fig. 3 D-gal-induced heart senescence is aggravated in miR-30a-5p KO mice. a Schematic of the experimental timeline of D-gal-induced aging in WT and KO mice. Representative ECG images (b) and analysis (c) of the LVEF and LVFS of Veh- or D-gal-induced WT and KO mice (n = 7 in each group). Representative immunofluorescence staining (d) and analysis (e) of γH2AX in heart tissues of Veh- or D-gal-induced WT and KO mice (n = 4 in each group, scale bars = 200 μm). f qRT‒PCR analysis of telomere length in heart tissues from Veh- or D-gal-induced WT and KO mice (n = 6 in each group). Representative western blotting images (g) and analysis (h) of p53 and p21 expression in heart tissues of Veh- or D- gal-induced WT and KO mice (n = 4 in each group). Representative WGA staining images (i) and analysis (j) of the hearts of Veh- or D-gal- induced WT and KO mice (n = 5 in each group; scale bars = 100 μm). k qRT‒PCR analysis of myocardial IL-1β (n = 3 in each group) and IL-6 (n = 4 in each group) levels in Veh- or D-gal-induced WT and KO mice. The data are presented as the means ± standard errors. Statistical significance was assessed using one-way ANOVA. ECG echocardiography, WT wild-type, KO knockout, LVEF left ventricular ejection fraction, LVFS left ventricular fractional shortening, D-gal D-galactose, qRT‒PCR quantitative real‒time polymerase chain reaction, γH2AX H2A histone family member X, Veh vehicle, WGA wheat germ agglutinin.

Article Snippet: The primary antibodies used for western blotting were mouse monoclonal anti-p53 (Cat# Ab26, Abcam, Cambridge, UK), rabbit polyclonal anti-p21 (Cat# 8248-1-AP) and mouse monoclonal anti-TERF2 (Cat# 66893- 1-Ig, both ProteinTech, Wuhan, China), rabbit polyclonal anti-TP53INP1 (Cat# OM204116, OmniMabs, Alhambra, CA, USA), mouse monoclonal antiSUMO1 (Cat# sc-5308, Santa Cruz, Dallas, Texas, USA), rabbit polyclonal anti-TERT (Cat# DF7129), rabbit polyclonal anti-DDIT4 (Cat# 10638-1-AP, Proteintech, Wuhan, China), and rabbit polyclonal anti-SENP1 (Cat# AF0275, both Affinity Bioscience, Nanjing, China).

Techniques: Staining, Western Blot, Expressing, Knock-Out, Polymerase Chain Reaction