Journal: bioRxiv

Article Title: The AMPKα2/PHF2 axis is critical for turning over lipid droplets during muscle stem cell fate

doi: 10.1101/2025.01.18.630727

Figure Lengend Snippet: CSA distribution of muscle fibers on cryosections of regenerated TA muscles at 11dpi (A) and 28dpi (B). (C) Representative images of immunostainings. (D) Number of myofibers per mm2 at 28dpi. (E) Quantification of the number of PAX7 pos Ki67 neg cells per mm 2 at 28 dpi. (F) Quantification of the number of MYOG pos cells per mm 2 on CTRL SC and PHF2 SCiKO TA cryosections at 11dpi. (G) Percentage of nuclei per myotube, (H) Percentage of PAX7 pos MYOG neg (yellow arrowhead) and (I) PAX7 neg MYOG pos cells (purple arrowhead) in cultures after 48h in LSM. (J) Representative images of immunostainings. (K) Experimental setup. Fusion index (L) and percentage of nuclei per myotube (M) in cultures after 48h in LSM. (N) Experimental setup. Percentage of nuclei per myotube (O) and of PAX7 pos cells (P) in PHF2 SCiKO cells transfected with either MOCK, PHF2 wt or PHF2 H249A . Scale bars, 50 μm. n = 3-6 mice/genotype. n = 3-8 primary cultures/genotype. Values are mean or percentage mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 (Student t -test [D-E-F-H-I-L panels], Sidak’s test after two-way ANOVA [A-B panels], Tukey’s test after one way [G-M-P panels] or two-way ANOVA [O panel]).

Article Snippet: They were transferred in a wet chamber for 1h30 at RT in 1% BSA,50 mM Tris-HCl, pH 8.2 for 1h30 at RT, labelled with gold conjugated Secondary antibody (Goat anti Rabbit 10nm for Myog and Goat anti Rabbit 5nm for Phf2) (Aurion).

Techniques: Muscles, Transfection

Journal: bioRxiv

Article Title: The AMPKα2/PHF2 axis is critical for turning over lipid droplets during muscle stem cell fate

doi: 10.1101/2025.01.18.630727

Figure Lengend Snippet: (A-B) Percentage of MYOG pos BODIPY pos cells transfected with either MOCK, PHF2 wt or PHF2 S655E and PHF2 S655A . Representative images. (C) Percentage of MYOG pos and (D) percentage of MYOG pos BODIPY pos cells after 48h in LSM. (E) Representative images. (F) Percentage of MYOG pos BODIPY pos cells transfected with either MOCK, PHF2 S655E or PHF2 S655A . Scale bars, 5 μm. n = 3 primary cultures/genotype. Values are mean or percentage mean ± SEM. ***P < 0.001 (Student t -test [C-D panels] and Tukey’s test after one way-ANOVA [B-F panels]).

Article Snippet: They were transferred in a wet chamber for 1h30 at RT in 1% BSA,50 mM Tris-HCl, pH 8.2 for 1h30 at RT, labelled with gold conjugated Secondary antibody (Goat anti Rabbit 10nm for Myog and Goat anti Rabbit 5nm for Phf2) (Aurion).

Techniques: Transfection

Journal: bioRxiv

Article Title: The AMPKα2/PHF2 axis is critical for turning over lipid droplets during muscle stem cell fate

doi: 10.1101/2025.01.18.630727

Figure Lengend Snippet: (A) Clustering of EYFP pos -FACS-isolated cells from CTRL SC and PHF2 SCiKO at 11dpi (n=3 mice pooled in 1 sample/genotype). (B) Myog pos myocytes cluster. Plin2, Plin3 (C) and Gdi2 (D) mRNA expression levels. (E) GDI2 protein level in CTRL SC and PHF2 SCiKO mononucleated cells 24h post LSM. (F) Integrative Genomics Viewer (IGV) snapshot depicting PHF2 enrichment (MACS peak call in dark blue) on the Gdi2 promoter region (GSM3462720). PLA analysis (G), electron microscopy analysis of LD-mitochondria contacts (H) and confocal analysis of LD-RAB8a-mitochondria contacts (I) in CTRL SC and PHF2 SCiKO mononucleated cells 24h post LSM. Scale bars, 1, 2 and 5 μm. n = 3 primary cultures/genotype. For (H panel), n=20 cells/genotype/conditions. Values are mean or percentage mean ± SEM. **P < 0.01, ****P < 0.0001 (Student t -test [E panel] and Tukey’s multiple comparison test after one way-ANOVA [G-H-I panels]).

Article Snippet: They were transferred in a wet chamber for 1h30 at RT in 1% BSA,50 mM Tris-HCl, pH 8.2 for 1h30 at RT, labelled with gold conjugated Secondary antibody (Goat anti Rabbit 10nm for Myog and Goat anti Rabbit 5nm for Phf2) (Aurion).

Techniques: Isolation, Expressing, Electron Microscopy, Comparison