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. 2021 Feb 2;10:e62174. doi: 10.7554/eLife.62174

Figure 2. The time course of ferritinophagy in the heart after transverse aortic constriction (TAC).

(A) Western blot analysis of FTH1 in Ncoa4+/+ and Ncoa4–/– hearts 4 weeks after TAC. GAPDH was used as the loading control. The right-hand graphs show the densitometric analysis. The average value for sham-operated Ncoa4+/+ hearts was set to 1 (biologically independent samples: n = 6). (B) mRNA expression of Fth1 in the heart 4 weeks after TAC. Gapdh mRNA was used as the loading control. The average value for sham-operated Ncoa4+/+ hearts was set to 1 (biologically independent samples: n = 8). (C–H) The Ncoa4+/+ mice were subjected to TAC and analyzed 3 days after sham operation and 3, 5, and 7 days and 4 weeks after the operation. (C) Heart homogenates after TAC were subjected to western blot analysis using anti-FTH1 antibody (n = 5 biologically independent samples for each group). GAPDH was used as the loading control. (D) Cardiac Fth1 mRNA levels after TAC (n = 5 biologically independent samples). Gapdh mRNA was used as the loading control. (E and F) Immunofluorescence analysis of LC3B (green) and FTH1 (red) in the heart after TAC (n = 5 biologically independent samples). (G and H) Immunofluorescence analysis of LAMP2a (green) and FTH1 (red) in the heart after TAC (n = 5 biologically independent samples). Scale bar, 5 μm in (E) and (G). Arrows indicate double-positive dots. The values are presented as the mean ± SEM. The data were evaluated by one-way analysis of variance (ANOVA), followed by Tukey–Kramer’s post hoc test. *p<0.05, **p<0.001, ***p<0.0001. NS, p>0.05 versus sham-operated group. Exact p-values are provided in Supplementary file 1.

Figure 2—source data 1. Source data for Figure 2.

Figure 2.

Figure 2—figure supplement 1. Echocardiographic parameter aftertransverse aortic constriction (TAC).

Figure 2—figure supplement 1.

Time course of changes in the echocardiographic parameters of left ventricular (LV) fractional shortening (FS), the end-diastolic LV internal dimension (LVIDd) and the end-systolic LV internal dimension (LVIDs). Echocardiographic analysis on Ncoa4+/+ and Ncoa4–/– mice were performed before (pre) and 3, 5, and 7 days after TAC (n = 3 biologically independent samples). The values are presented as the mean ± SEM. Two-way analysis of variance (ANOVA) followed by Tukey's multiple comparisons test was used. *p<0.05, **p<0.001, ***p<0.0001. p>0.05, sham-operated Ncoa4+/+ versus TAC-operated Ncoa4+/+. †p<0.05, ††p<0.001, †††p<0.0001. p>0.05, sham-operated Ncoa4–/– versus TAC-operated Ncoa4–/–. ‡p<0.05, ‡‡p<0.001, ‡‡‡p<0.0001. p>0.05, TAC-operated Ncoa4+/+ versus TAC-operated Ncoa4–/–. Exact p-values are provided in Supplementary file 1.
Figure 2—figure supplement 1—source data 1. Source data for Figure 2—figure supplement 1.