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. Author manuscript; available in PMC: 2023 Dec 20.
Published in final edited form as: Circulation. 2022 Nov 24;146(25):1950–1967. doi: 10.1161/CIRCULATIONAHA.122.061960

Figure 4. CM-specific HMGCS2 knockout mice present a metabolic switch along with impaired mitochondria leading to a pathological hypertrophy.

Figure 4.

A, Schematic diagram showing establishment of CM-specific HMGCS2 knockout mice. B, RNA and protein expression of HMGCS2 detected by real-time PCR and western-blot in isolated CMs from 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− mice. 40 μg protein was loaded for examination. RNA expression was normalized by Gapdh. C, OHB level measured by OHB colorimetric assay in isolated CMs from 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− mice. D, Ejection fraction of 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− hearts measured by echocardiography. E, Left ventricular mass of 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− hearts measured by echocardiography. F, RNA expression of hypertrophic marker βMHC and αMHC measured by real-time PCR in the isolated CMs from 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− mice. RNA expression was normalized by Gapdh. G, RNA expression of PGC1α measured by real-time PCR in the isolated CMs from 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− mice. H, The fibrotic area in 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− hearts shown by Masson Trichrome staining of heart tissue sections. I, Quantification of fibrotic percentage in 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− hearts measured by Masson Trichrome Staining. J, Quantification of CM size in 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− hearts measured by Masson Trichrome Staining. K, Immunofluorescence of heart tissue sections to present morphology of proliferative CMs through H3P and cTnT staining on 1-day-old control or CM-HMGCS2−/− hearts. Arrow Heads represented H3P+/cTnT+ proliferative CMs. Scale bars were 50 μm. L, Quantification of proliferative CM percentage (H3P+%) in the heart tissue sections of 1-day-old control or CM-HMGCS2−/− hearts. M, Immunofluorescence of heart tissue sections showing morphology of proliferative CMs through AURKB and cTnT staining on 1-day-old control or CM-HMGCS2−/− hearts. Arrow Heads represented AURKB+/cTnT+ proliferative CMs. Scale bars were 50 μm. N, The statistics of proliferative CM percentage (AURKB+%) in the heart tissue sections of 1-day-old control or CM-HMGCS2−/− hearts. O, OCR detected by Seahorse analysis in the isolated CMs from CM-HMGCS2+/+ or CM-HMGCS2−/− mice. P, Quantification of basal and maximal OCRs in the isolated CMs from 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− mice. Q, Mitochondrial copy numbers detected by mtDNA through real-time PCR in the isolated CMs from 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− mice. R, Mitochondrial RNA expression detected by real-time PCR in adult CMs isolated from 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− mice. RNA expressions were normalized by Gapdh. S, Mitochondrial structure examined by TEM in 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− hearts. T, Mitochondrial size of isolated 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− hearts in TEM data. U, The aspect ratio of mitochondrial length-to-width in isolated 10-week-old CM-HMGCS2+/+ or CM-HMGCS2−/− hearts in TEM data.