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( A ) Illustration of the study design of <t>CoQ10</t> treatment in mice. ( B ) Concentration of CoQ10 in TA tissues of mice at 2 weeks ( n = 3 mice). P value: ** P = 0.0013; ns- P = 0.4524; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, * P = 0.0118; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10, * P = 0.0435. ( C ) Quantification of ATP content in TA tissues from Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks, with or without CoQ10 treatment ( n = 3 mice). Values were normalized to total protein levels. P value: ** P = 0.0012; * P = 0.0119; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, ns- P = 0.9377; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10, ns- P = 0.2180. ( D ) Representative TEM images of TA sections from Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks. Black arrows indicate damaged mitochondria. The percentage of damaged mitochondria was quantified using ImageJ software ( n = 8 areas). Scale bars: 1 µm. P value: Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+Vehicle ** P = 0.0017; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, ** P = 0.0066; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10 ** P = 0.0022; Foxk2 fl/fl ; Myod1 -Cre+Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+ CoQ10, ** P = 0.0057. ( E ) ROS production in TA sections of Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks, with or without CoQ10 treatment, measured by DHE staining ( n = 4 areas). Quantifications were performed using ImageJ software. Scare bar: 200 μm. P value: Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+Vehicle * P = 0.0325; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, ns- P = 0.1736; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10 ns- P = 0.2200 ; Foxk2 fl/fl ; Myod1 -Cre+Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+ CoQ10, * P = 0.0426. ( F ) H&E staining illustrating histological aspects and the percentage of central nuclei myofibers in skeletal muscles (Gas, TA, IM, TM, Dp, and EM) of Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks, with or without CoQ10 treatment ( n = 4 areas). Black arrows indicate central nuclei. The inset in the upper right corner (scale bars: 25 µm) is an enlarged view of the boxed area in the main image, highlighting central nuclei myofibers (scale bars: 100 µm). P value for Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, Foxk2 fl/fl ; Myod1 -Cre+Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10, Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+Vehicle and Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10: Gas: ns- P = 0.6529; * P = 0.0225; *** P = 0.0003; *** P = 0.0007. TA: ns- P = 0.4640; ns- P = 0.2498; *** P = 0.0008; ** P = 0.0011. IM: ns- P = 0.5588; ** P = 0.0067; *** P = 0.0004; * P = 0.0337. TM: ns- P = 0.1669; * P = 0.0110; ** P = 0.0044; ns- P = 0.1219. Dp: ns- P = 0.2431; * P = 0.0322; ** P = 0.0077; ns- P = 0.6149. EM: ns- P = 0.6625; ** P = 0.0024; *** P = 0.0001; ns- P = 0.4264. ( G ) IF staining of Laminin in the TA of Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks with or without CoQ10 treatment. The distribution of CSA of myofibers was calculated ( n = 3 areas). Scale bars: 100 µm. P value: 500-899, ** P = 0.0063, * P = 0.0198; 900-1299, ** P = 0.0010; 1300-1699, * P = 0.03871; 1700-2099, * P = 0.0251; 2100-2499, ** P = 0.0022; >2500, ** P = 0.0060. Data were analyzed by Student’s t test. All error bars indicate mean ± standard deviation. CoQ10 Coenzyme Q10, Gas gastrocnemius, TA tibialis anterior, IM intercostal muscle, TM tongue muscle, Dp diaphragm, EM eyelid muscle, TEM transmission electron microscopy, ROS reactive oxygen species, DHE dihydroethidium, H&E hematoxylin-eosin staining, IF immunofluorescence, CSA cross-sectional area.
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( A ) Illustration of the study design of CoQ10 treatment in mice. ( B ) Concentration of CoQ10 in TA tissues of mice at 2 weeks ( n = 3 mice). P value: ** P = 0.0013; ns- P = 0.4524; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, * P = 0.0118; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10, * P = 0.0435. ( C ) Quantification of ATP content in TA tissues from Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks, with or without CoQ10 treatment ( n = 3 mice). Values were normalized to total protein levels. P value: ** P = 0.0012; * P = 0.0119; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, ns- P = 0.9377; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10, ns- P = 0.2180. ( D ) Representative TEM images of TA sections from Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks. Black arrows indicate damaged mitochondria. The percentage of damaged mitochondria was quantified using ImageJ software ( n = 8 areas). Scale bars: 1 µm. P value: Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+Vehicle ** P = 0.0017; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, ** P = 0.0066; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10 ** P = 0.0022; Foxk2 fl/fl ; Myod1 -Cre+Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+ CoQ10, ** P = 0.0057. ( E ) ROS production in TA sections of Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks, with or without CoQ10 treatment, measured by DHE staining ( n = 4 areas). Quantifications were performed using ImageJ software. Scare bar: 200 μm. P value: Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+Vehicle * P = 0.0325; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, ns- P = 0.1736; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10 ns- P = 0.2200 ; Foxk2 fl/fl ; Myod1 -Cre+Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+ CoQ10, * P = 0.0426. ( F ) H&E staining illustrating histological aspects and the percentage of central nuclei myofibers in skeletal muscles (Gas, TA, IM, TM, Dp, and EM) of Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks, with or without CoQ10 treatment ( n = 4 areas). Black arrows indicate central nuclei. The inset in the upper right corner (scale bars: 25 µm) is an enlarged view of the boxed area in the main image, highlighting central nuclei myofibers (scale bars: 100 µm). P value for Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, Foxk2 fl/fl ; Myod1 -Cre+Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10, Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+Vehicle and Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10: Gas: ns- P = 0.6529; * P = 0.0225; *** P = 0.0003; *** P = 0.0007. TA: ns- P = 0.4640; ns- P = 0.2498; *** P = 0.0008; ** P = 0.0011. IM: ns- P = 0.5588; ** P = 0.0067; *** P = 0.0004; * P = 0.0337. TM: ns- P = 0.1669; * P = 0.0110; ** P = 0.0044; ns- P = 0.1219. Dp: ns- P = 0.2431; * P = 0.0322; ** P = 0.0077; ns- P = 0.6149. EM: ns- P = 0.6625; ** P = 0.0024; *** P = 0.0001; ns- P = 0.4264. ( G ) IF staining of Laminin in the TA of Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks with or without CoQ10 treatment. The distribution of CSA of myofibers was calculated ( n = 3 areas). Scale bars: 100 µm. P value: 500-899, ** P = 0.0063, * P = 0.0198; 900-1299, ** P = 0.0010; 1300-1699, * P = 0.03871; 1700-2099, * P = 0.0251; 2100-2499, ** P = 0.0022; >2500, ** P = 0.0060. Data were analyzed by Student’s t test. All error bars indicate mean ± standard deviation. CoQ10 Coenzyme Q10, Gas gastrocnemius, TA tibialis anterior, IM intercostal muscle, TM tongue muscle, Dp diaphragm, EM eyelid muscle, TEM transmission electron microscopy, ROS reactive oxygen species, DHE dihydroethidium, H&E hematoxylin-eosin staining, IF immunofluorescence, CSA cross-sectional area.

Journal: EMBO Molecular Medicine

Article Title: FOXK2 in skeletal muscle development: a new pathogenic gene for congenital myopathy with ptosis

doi: 10.1038/s44321-025-00247-x

Figure Lengend Snippet: ( A ) Illustration of the study design of CoQ10 treatment in mice. ( B ) Concentration of CoQ10 in TA tissues of mice at 2 weeks ( n = 3 mice). P value: ** P = 0.0013; ns- P = 0.4524; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, * P = 0.0118; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10, * P = 0.0435. ( C ) Quantification of ATP content in TA tissues from Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks, with or without CoQ10 treatment ( n = 3 mice). Values were normalized to total protein levels. P value: ** P = 0.0012; * P = 0.0119; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, ns- P = 0.9377; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10, ns- P = 0.2180. ( D ) Representative TEM images of TA sections from Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks. Black arrows indicate damaged mitochondria. The percentage of damaged mitochondria was quantified using ImageJ software ( n = 8 areas). Scale bars: 1 µm. P value: Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+Vehicle ** P = 0.0017; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, ** P = 0.0066; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10 ** P = 0.0022; Foxk2 fl/fl ; Myod1 -Cre+Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+ CoQ10, ** P = 0.0057. ( E ) ROS production in TA sections of Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks, with or without CoQ10 treatment, measured by DHE staining ( n = 4 areas). Quantifications were performed using ImageJ software. Scare bar: 200 μm. P value: Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+Vehicle * P = 0.0325; Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, ns- P = 0.1736; Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10 ns- P = 0.2200 ; Foxk2 fl/fl ; Myod1 -Cre+Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+ CoQ10, * P = 0.0426. ( F ) H&E staining illustrating histological aspects and the percentage of central nuclei myofibers in skeletal muscles (Gas, TA, IM, TM, Dp, and EM) of Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks, with or without CoQ10 treatment ( n = 4 areas). Black arrows indicate central nuclei. The inset in the upper right corner (scale bars: 25 µm) is an enlarged view of the boxed area in the main image, highlighting central nuclei myofibers (scale bars: 100 µm). P value for Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl + CoQ10, Foxk2 fl/fl ; Myod1 -Cre+Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10, Foxk2 fl/fl +Vehicle vs Foxk2 fl/fl ; Myod1 -Cre+Vehicle and Foxk2 fl/fl + CoQ10 vs Foxk2 fl/fl ; Myod1 -Cre+CoQ10: Gas: ns- P = 0.6529; * P = 0.0225; *** P = 0.0003; *** P = 0.0007. TA: ns- P = 0.4640; ns- P = 0.2498; *** P = 0.0008; ** P = 0.0011. IM: ns- P = 0.5588; ** P = 0.0067; *** P = 0.0004; * P = 0.0337. TM: ns- P = 0.1669; * P = 0.0110; ** P = 0.0044; ns- P = 0.1219. Dp: ns- P = 0.2431; * P = 0.0322; ** P = 0.0077; ns- P = 0.6149. EM: ns- P = 0.6625; ** P = 0.0024; *** P = 0.0001; ns- P = 0.4264. ( G ) IF staining of Laminin in the TA of Foxk2 fl/fl and Foxk2 fl/fl ; Myod1 -Cre littermates at 2 weeks with or without CoQ10 treatment. The distribution of CSA of myofibers was calculated ( n = 3 areas). Scale bars: 100 µm. P value: 500-899, ** P = 0.0063, * P = 0.0198; 900-1299, ** P = 0.0010; 1300-1699, * P = 0.03871; 1700-2099, * P = 0.0251; 2100-2499, ** P = 0.0022; >2500, ** P = 0.0060. Data were analyzed by Student’s t test. All error bars indicate mean ± standard deviation. CoQ10 Coenzyme Q10, Gas gastrocnemius, TA tibialis anterior, IM intercostal muscle, TM tongue muscle, Dp diaphragm, EM eyelid muscle, TEM transmission electron microscopy, ROS reactive oxygen species, DHE dihydroethidium, H&E hematoxylin-eosin staining, IF immunofluorescence, CSA cross-sectional area.

Article Snippet: In the mouse study, we selected an optimal administration method for CoQ10 (MCE) supplementation, delivering 600 mg/kg/day (dissolved in corn oil) via oral gavage, as previously described (Pan et al, ).

Techniques: Concentration Assay, Software, Staining, Muscles, Standard Deviation, Transmission Assay, Electron Microscopy, Immunofluorescence