Journal: Cell stem cell

Article Title: The mitochondrial protein OPA1 regulates the quiescent state of adult muscle stem cells

doi: 10.1016/j.stem.2022.07.010

Figure Lengend Snippet: Key Resource Table

Article Snippet: Single EDL myofibers were treated with 2 μM Mdivi-1 (Millipore Sigma; M0199), 10 μM MitoTEMPO (Millipore Sigma; SML0737), 10 or 50 μM BSO (Millipore Sigma; B2515), 10 nM (low dose) or 50 μM Rotenone (Millipore Sigma; R8875), 25nM (low dose) mito-Paraquat (Abcam, ab146819), 1 mM L-Glutathione (GSH) (Millipore Sigma; G4251), 5 mM L-Cysteine (Millipore Sigma; 5360), 5 mM Glycine (Bio-Rad; 1610717), 50 μM Antimycin A (Millipore Sigma; A8674), 5 μM Oligomycin (Millipore Sigma; O4876), 20 nM Rapamycin (Invivogen; tlrl-rap), 10 nM Torin-1 (Invivogen; inh-tor1), 40 ng/mL Recombinant Mouse HGF-A (R&D Systems; 1200-SE) or 10 ng/mL Recombinant human active HGF (Abcam; Ab632), depending on experimental design.

Techniques: Recombinant, TUNEL Assay, Isolation, Enzyme-linked Immunosorbent Assay, Microarray, Software, Microscopy, Imaging

Journal: Cell stem cell

Article Title: The mitochondrial protein OPA1 regulates the quiescent state of adult muscle stem cells

doi: 10.1016/j.stem.2022.07.010

Figure Lengend Snippet: Key Resource Table

Article Snippet: Single EDL myofibers were treated with 2 μM Mdivi-1 (Millipore Sigma; M0199), 10 μM MitoTEMPO (Millipore Sigma; SML0737), 10 or 50 μM BSO (Millipore Sigma; B2515), 10 nM (low dose) or 50 μM Rotenone (Millipore Sigma; R8875), 25nM (low dose) mito-Paraquat (Abcam, ab146819), 1 mM L-Glutathione (GSH) (Millipore Sigma; G4251), 5 mM L-Cysteine (Millipore Sigma; 5360), 5 mM Glycine (Bio-Rad; 1610717), 50 μM Antimycin A (Millipore Sigma; A8674), 5 μM Oligomycin (Millipore Sigma; O4876), 20 nM Rapamycin (Invivogen; tlrl-rap), 10 nM Torin-1 (Invivogen; inh-tor1), 40 ng/mL Recombinant Mouse HGF-A (R&D Systems; 1200-SE) or 10 ng/mL Recombinant human active HGF (Abcam; Ab632), depending on experimental design.

Techniques: Recombinant, TUNEL Assay, Isolation, Enzyme-linked Immunosorbent Assay, Microarray, Software, Microscopy, Imaging

Journal: Cell stem cell

Article Title: The mitochondrial protein OPA1 regulates the quiescent state of adult muscle stem cells

doi: 10.1016/j.stem.2022.07.010

Figure Lengend Snippet:

Article Snippet: Single EDL myofibers were treated with 2 μM Mdivi-1 (Millipore Sigma; M0199), 10 μM MitoTEMPO (Millipore Sigma; SML0737), 10 or 50 μM BSO (Millipore Sigma; B2515), 10 nM (low dose) or 50 μM Rotenone (Millipore Sigma; R8875), 25nM (low dose) mito-Paraquat (Abcam, ab146819), 1 mM L-Glutathione (GSH) (Millipore Sigma; G4251), 5 mM L-Cysteine (Millipore Sigma; 5360), 5 mM Glycine (Bio-Rad; 1610717), 50 μM Antimycin A (Millipore Sigma; A8674), 5 μM Oligomycin (Millipore Sigma; O4876), 20 nM Rapamycin (Invivogen; tlrl-rap), 10 nM Torin-1 (Invivogen; inh-tor1), 40 ng/mL Recombinant Mouse HGF-A (R&D Systems; 1200-SE) or 10 ng/mL Recombinant human active HGF (Abcam; Ab632), depending on experimental design.

Techniques: Concentration Assay, Recombinant

Journal: Science advances

Article Title: Mg 2+ influx mediated by TRPM7 triggers the initiation of muscle stem cell activation.

doi: 10.1126/sciadv.adu0601

Figure Lengend Snippet: Fig. 5. Loss of TRPM7 impairs activation of the mTOR signaling and induction of GAlert in MuSCs. (A) Time course for induction of Trpm7 deficiency and isolation of myofibers. (B and C) Analysis of MuSCs on myofibers cultured for 30 h. (B) MuSC size: Representative YFP-positive MuSC images and size distribution (>100 cells per condi- tion). (C) pS6: Representative fluorescent images and quantification of pS6 intensity (>50 cells per condition, N = 3 mice). (D) Time course for induction of Trpm7 defi- ciency, injection of CTX, and isolation of myofibers. MuSCs on myofibers without injury was defined as quiescent satellite cells (QSCs). MuSCs on myofibers isolated from the contralateral leg of injury was defined as contralateral satellite cells (CSCs). Myofibers were isolated 4 days after CTX injection for GAlert induction. (E) Morphological evaluation of MuSCs after induction of GAlert. Phase contrast images of con (top) and cKO (bottom) MuSCs. Size distribution of QSCs and CSCs (>100 MuSCs per condition; data are individually plotted.). (F) pS6 in QSCs and CSCs: Representative images and quantification in con and cKO MuSCs (>50 cells per condition, N > 3 mice). (G) FACS histograms of MitoTracker and PyroninY staining. Gray: Unstained MuSCs; blue: con QSCs; green: con CSCs; red: cKO CSCs. (H) Time course for induction of Trpm7 defi- ciency, injection of HGFA, and isolation of myofibers. MuSCs from HGFA-treated mice were defined as HGFA for GAlert induction. PBS was used for control. (I) HGFA effect on pS6 in cKO MuSCs: Representative images and quantification of pS6 in con and cKO MuSCs after GAlert induction with HGFA (>50 cells per condition, N = 3 mice). Scale bars, 10 μm in [(B), (C), (E), (F), and (I)].

Article Snippet: Recombinant HGFA (#1200- SE, R&D Systems) was administered to mice at a dose of 1 μg diluted in 200 μl of PBS via tail vein injection as previously described (42).

Techniques: Activation Assay, Isolation, Cell Culture, Injection, Staining, Control

Journal: International journal of molecular sciences

Article Title: Combined Therapy Targeting MET and Pro-HGF Activation Shows Significant Therapeutic Effect Against Liver Metastasis of CRPC.

doi: 10.3390/ijms26052308

Figure Lengend Snippet: Figure 3. Inhibition of MET and HGF-activation in CRTC2 cells, in vitro analysis. (A) CRTC2 cells were pre-cultured in FBS-free DMEM for 24 h. The cells were then treated with each agent (final concentration of 250 nM for MET-I and 10 µM for HGFA-I) at 37 ◦C for 2 h, followed by the addition of mouse recombinant pro-HGF (50 ng/mL). After incubation at 37 ◦C for 2 h, the proteins were extracted. Phosphorylation of MET and expression of total-MET, HGF, and α-tubulin were determined by immunoblot analysis. (B) CRTC2 cells were pre-cultured in FBS-free DMEM for 24 h. The cells were seeded in a 96-well plate at a density of 1 × 104 cells/100 µL in FBS-free DMEM. The cells were then treated with each agent (final concentration of 20 µM for MET-I and 20 µM for HGFA-I) at 37 ◦C for 2 h, followed by the addition of 50 ng/mL mouse recombinant pro-HGF, and incubated at 37 ◦C for 48 h. Cell proliferation was analyzed by detecting the absorbance at 490 nm and calculated as a ratio to the absorbance at day 0 (before administration, ** p < 0.01).

Article Snippet: Recombinant mouse HGF propeptide protein (#7058-HG) and mouse HGF protein (#2207-HG) were purchased from R&D Systems.

Techniques: Inhibition, Activation Assay, In Vitro, Cell Culture, Concentration Assay, Recombinant, Incubation, Phospho-proteomics, Expressing, Western Blot