Journal: EMBO Reports

Article Title: The muscle specific MEF2Dα2 isoform promotes muscle ketolysis and running capacity in mice

doi: 10.1038/s44319-025-00578-3

Figure Lengend Snippet: ( A ) RT-PCR analysis of the mutually exclusive α-exons in Mef2d transcript using total RNA from the indicated fetal hind limb muscle and tissues from adult wild-type mice. ( B ) RT-PCR analysis of Mef2d α exons using soleus RNA from line 2. The numbers indicate PSI; data are mean ± SD; n = 3. The numbers indicate the percent spliced in (PSI). Bolded numbers are significant by Student T test. ( C ) RT-PCR analysis of the alternative exons in Mef2c and Mef2a transcripts using total RNA from gastrocnemius muscle from line 1. Data are mean ± SD; n = 3. PSI for exon is indicated and was not significantly different between the genotypes by Student’s t test. ( D ) Western blot showing MEF2D protein levels in TA (upper left) and Soleus muscles (upper right) from line 2. The bottom panels show Coomassie stained blots, from the upper panels showing total protein loaded. The numbers are mean ± SD; n = 3. Student’s t test found no differences in genotypes. ( E ) RT-qPCR showing relative mRNA levels Mef2d , Mef2a , and Mef2c relative to Rpl30 using total RNA in the indicated muscle groups from line 2 mice. Data are mean ± SEM; n = 4. ** P = 0.0088 (multiple Student’s t test, unpaired). .

Article Snippet: MEF2D (ChIP) , Proteintech , 14353-1-AP.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Muscles, Staining, Quantitative RT-PCR

Journal: EMBO Reports

Article Title: The muscle specific MEF2Dα2 isoform promotes muscle ketolysis and running capacity in mice

doi: 10.1038/s44319-025-00578-3

Figure Lengend Snippet: ( A ) RT-PCR analysis of the mutually exclusive α-exons and alternative β exon in Mef2d transcript using total RNA from the indicated muscle groups from Wt and Mef2dα2 Eko mice from line 1. The numbers indicate the percent spliced in (PSI) for indicated exon. Data are mean ± SD; n = 3. Bold numbers indicate significance by Student t test. ( B ) Western blot showing MEF2D protein levels in TA (upper left) and Soleus muscles (upper right) from line 1. Bottom panels show Coomassie stained blots from the upper panels showing total protein loaded. The numbers indicate the relative protein level normalized to total protein by Coomassie stained blots. Data are mean ± SD; n = 3. ( C ) RT-qPCR showing mRNA levels Mef2d , Mef2a , and Mef2c relative to Rpl30 using total RNA in the indicated muscle groups from line 1 mice. Data are mean ± SEM; n = 3. No significant changes were found between the genotypes by multiple t test in ( B , C ). .

Article Snippet: MEF2D (ChIP) , Proteintech , 14353-1-AP.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Muscles, Staining, Quantitative RT-PCR

Journal: EMBO Reports

Article Title: The muscle specific MEF2Dα2 isoform promotes muscle ketolysis and running capacity in mice

doi: 10.1038/s44319-025-00578-3

Figure Lengend Snippet: ( A ) RNA-sequencing data using poly-A selected RNA from WT and MEF2Dα2 Eko quadriceps muscles were aligned to mm10 UCSC browser showing Mef2d α exons region. ( B ) Graph showing number of genes that were down or upregulated in MEF2Dα2 Eko muscles in comparison to age and sex-matched WT muscles (FDR < 0.05). ( C ) RT-qPCR showing relative expression of Bdh1, Oxct1, and Acat1 transcripts normalized to Hprt transcript levels in WT and MEF2Dα2 Eko soleus muscles from line 1. Data are mean ± SEM; n ≥ 6. ** P < 0.01, *** P < 0.001 (Student’s t test). The exact P value is indicated above the bar graphs. ( D ) Western blot showing BDH1 level in soleus muscles of indicated mice from line 1. The panel on the right show BDH1, OXCT1, and ACAT1 level when normalized to total protein loaded as estimated by Coomassie staining of the same blot. Data are mean ± SEM, n = 3, * P < 0.016, ** P < 0.0042, **** P = 0. 000082 (Student’s t test). .

Article Snippet: MEF2D (ChIP) , Proteintech , 14353-1-AP.

Techniques: RNA Sequencing, Muscles, Comparison, Quantitative RT-PCR, Expressing, Western Blot, Staining

Journal: EMBO Reports

Article Title: The muscle specific MEF2Dα2 isoform promotes muscle ketolysis and running capacity in mice

doi: 10.1038/s44319-025-00578-3

Figure Lengend Snippet: ( A ) β-hydroxybutyrate (BHB) tolerance test in untrained 28-week-old male sedentary mice. Graphs displays mean ± SEM, with n ≥ 9. * P < 0.05 ( P = 0.02525 at 45 min, P = 0.021921 at 60 min), ** P < 0.01 ( P = 0.002777 at 90 min) by multiple t test between genotypes at different time-points. ( B ) The indicated age-matched male mice were run for 55 min at 18 m/min, and BHB was measured immediately after running (0 h), and 1–3 h post-run. The pre-run BHB values are from the same mice a day before the experiment. Data are mean ± SD, n = 10, n.s. not significant; P > 0.05, ** P < 0.01 ( P = 0.00121 at 0 h), * P < 0.05 ( P = 0.0485 at 2-h post-exercise), *** P < 0.001 (0.000233088 at 3-h post-exercise) by multi p le t test between genotypes at different time-points. ( C ) The indicated age-matched male mice were fed control or ketogenic diet for 2-weeks and BHB was measured without fasting. Data are mean ± SD; n ≥ 8. The exact P value as calculated by one-way ANOVA is indicated above the bar graphs. ( D ) Comparison between state 3 JO 2 (ADP 200 µM) for isolated mitochondria from WT and MEF2Dα2 Eko skeletal muscles in presence of alpha-ketoglutarate (AKG) and AKG+Acetoacetate (AcAc). The representative time courses for these experiments are shown in Fig. . Data are mean ± SD; n = 5. The exact P value as calculated by one-way ANOVA is indicated above the bar graphs. ( E ) ChIP quantitative PCR for indicated genes using MEF2D and Serotype control (IgG) are shown (left), representative DNA product visualized on a 5% acrylamide gel (right). Data are mean ± SD; n = 4. For Bdh1, actual P value is indicated; for others, * P < 0.05, ** P < 0.01 (paired Student’s t test). The exact P value is indicated above the bar graphs. .

Article Snippet: MEF2D (ChIP) , Proteintech , 14353-1-AP.

Techniques: Control, Comparison, Isolation, Muscles, Real-time Polymerase Chain Reaction, Acrylamide Gel Assay

Journal: EMBO Reports

Article Title: The muscle specific MEF2Dα2 isoform promotes muscle ketolysis and running capacity in mice

doi: 10.1038/s44319-025-00578-3

Figure Lengend Snippet: ( A ) RT-qPCR showing relative expression of indicated transcripts normalized to Hprt transcript levels using total RNA from WT and MEF2Dα2 Eko mice livers. Data are mean ± SEM, n = 5, Student’s t test found no differences in genotypes. ( B ) Representative time-courses of isolated mitochondrial respiration for WT and MEF2Dα2 Eko mice transitioning from state 1 to state 4 respiration under AKG± AcAc. The respiratory rates are expressed as nmol/min/mg mitochondrial protein. The transitions from state 1 to state 4 respiration were monitored by first adding isolated mitochondria (0.05 mg/mL) to the respiration buffer at t = 0 min leading to state 1. At t = 2 min, substrates were added to energize the mitochondria, which led to state 2 respiration. This was followed by sequential additions of incremental ADP concentrations (100 and 200 µM). AKG: Alpha-ketoglutarate and AcAc: acetoacetate. ( C ) MEF2D ChIP data from Gönczi et al viewed on IGV viewer (version 2.19.4) for Igfn1, Bdh1, Oxct1, and Acat1 gene locus. The bi-directional arrowed line shows the region where we designed our primers for our analyses. .

Article Snippet: MEF2D (ChIP) , Proteintech , 14353-1-AP.

Techniques: Quantitative RT-PCR, Expressing, Isolation