primary antibodies of anti-mef2c  (Millipore)

Journal: Nature Cardiovascular Research

Article Title: Oxidative phosphorylation is required for cardiomyocyte re-differentiation and long-term fish heart regeneration

doi: 10.1038/s44161-025-00718-x

Figure Lengend Snippet: a , Representative images of immunofluorescence staining showing Mef2 + PCNA + double-positive proliferating cardiomyocytes in TU and WIK at 7 dpci. Framed areas highlight the wound border zone (red) (scale bar, 300 μm). b , Quantification of Mef2 + PCNA + double-positive cells showing differences in proliferating cardiomyocytes in the border zone at 7 dpci but not at 21 dpci. c , Positive correlation of percentage of proliferating cardiomyocytes in the border zone between 7 dpci and 21 dpci. d , No correlation between 7 dpci border zone proliferation and 90 dpci wound length. e , Venn diagram displaying complete lack of overlap between genes correlating to 7 dpci border zone proliferation and 90-dpci wound length. f , g , No correlation between border zone proliferation and OXPHOS ( f ) or Glycolysis ( g ) at 7 dpci. h , Quantification of Mef2 + PCNA + cells showing no difference in proliferating cardiomyocytes in the border zone of 7 dpci KCL adult treated with inhibitor PF-04859989 or rotenone compared to DMSO control. i , Temporal wound length reduction of all strains between 1, 7, 21 and 90 dpci. Arrow highlighting the strong decrease in wound length in WIK between 7 dcpi and 21 dpci. j , Percentage of hearts completely regenerated at 90 dpci or with closed compact wall but remaining internal scar or with open compact wall and internal scar remaining. b , 7 dpci: AB, NA, TU n = 7; SAT, WIK n = 6; TL n = 5; KCL n = 3, 21 dpci: AB, SAT, TL, TU n = 5; NA n = 6; WIK n = 7; KCL n = 8 (biological replicates); h , PF-04859989, DMSO n = 6, rotenone n = 5 (biological replicates); i , AB: 1, 90 dpci n = 7; 7 dpci n = 8; 21 dpci n = 6. NA: 1, 7, 90 dpci n = 7; 21 dpci n = 11. SAT: 1, 7, 21, 90 dpci n = 7. TL: 1, 7, 90 dpci n = 7; 21 dpci n = 9. TU: 1, 7 dpci n = 7; 21 dpci n = 5; 90 dpci n = 6. WIK: 1, 7, 21, 90 dpci n = 7. KCL: 1, 7, 21, 90 dpci n = 8 (biological replicates); j , AB, NA, SAT, TL, WIK n = 7; TU n = 6; KCL n = 8. b , h , One-way ANOVA with Tukey’s test. c , d , f , g , Simple linear regression. h , Two-way ANOVA with Tukey’s test. i , Data presented as mean ± s.e.m. Mef2, myocyte enhancer factor 2; PCNA, proliferating cell nuclear antigen.

Article Snippet: Primary antibodies Mef2c (Biorbyt, orb256682), PCNA (clone PC10; Dako, M0879), GFP (Abcam, ab13970), MF20 (Developmental Studies Hybridoma Bank (DSHB), AB_2147781) and embcmhc N2.261 (DSHB, AB_531790) and secondary antibodies Alexa Fluor 488 (Invitrogen, anti-mouse A11001, anti-chick A11039 and anti-rabbit A21206) and Alexa Fluor 555 (Invitrogen, A31570 ) were prepared using TNB buffer at a ratio of 1:200.

Techniques: Immunofluorescence, Staining, Control

Journal: Aging (Albany NY)

Article Title: lncRNA GPRC5D-AS1 as a ceRNA inhibits skeletal muscle aging by regulating miR-520d-5p

doi: 10.18632/aging.205279

Figure Lengend Snippet: The effect of overexpression of GPRC5D-AS1 on muscle regulatory factors. ( A ) qRT-PCR analyzed gene expression of Myf5, MyoG, MyoD and Mef2c. HSMM was control group. Dex (15 mM) was added in HSMM to establish atrophy cell model (model group). Empty plasmid (NC group) and GPRC5D-AS1-OE plasmid (lncRNA-OE group) were transfected into atrophy cell model and incubated for 48 h. * P < 0.05, ** P < 0.01 compared with control group; # P < 0.05, ## P < 0.01 compared with model group; & P < 0.05, && P < 0.01 compared with NC group. ( B ) Protein expression of Myf5, MyoG, MyoD and Mef2c detected by Western blot. Groups were set as previously mentioned. Empty plasmid and GPRC5D-AS1-OE plasmid were transfected into atrophy cell model and incubated for 48 h. ( C ) Quantitative analysis of western blot. * P < 0.05, ** P < 0.01 compared with control group; # P < 0.05, ## P < 0.01 compared with model group; & P < 0.05, && P < 0.01 compared with NC group.

Article Snippet: Then, protein was transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, USA), blocked with 5% skim milk and incubated with primary antibodies of anti-Mef2c (Cal.

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Plasmid Preparation, Transfection, Incubation, Western Blot

Journal: Aging (Albany NY)

Article Title: lncRNA GPRC5D-AS1 as a ceRNA inhibits skeletal muscle aging by regulating miR-520d-5p

doi: 10.18632/aging.205279

Figure Lengend Snippet: LncRNA GPRC5D-AS1 interacted with miR-520d-5p to promote myoblast proliferation. ( A , B ) qRT-PCR analyzed gene expression of GPRC5D-AS1, miR-520d-5p, MyoD1, MyoG, Mef2c, Myf5 and Wnt5a. 15 mM Dex was added in human skeletal muscle myoblasts to establish atrophy cell model (control group). Empty plasmid (NC group), GPRC5D-AS1-OE (lncRNA-OE group), GPRC5D-AS1-OE + miR-520d-5p mimic (lncRNA-OE + mimic group), miRNA inhibitor control (inhibitor NC group), miR-520d-5p inhibitor (miRNA inhibitor group) or MYOD1-OE plasmid (mRNA-OE group) was transfected into atrophy cell model and incubated for 48 h. * P < 0.05, ** P < 0.01 compared with control group; # P < 0.05, ## P < 0.01 compared with NC group; & P < 0.05, && P < 0.01 compared with lncRNA-OE group; % P < 0.05, %% P < 0.01 compared with inhibitor NC group.

Article Snippet: Then, protein was transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, USA), blocked with 5% skim milk and incubated with primary antibodies of anti-Mef2c (Cal.

Techniques: Quantitative RT-PCR, Expressing, Plasmid Preparation, Transfection, Incubation

Journal: Aging (Albany NY)

Article Title: lncRNA GPRC5D-AS1 as a ceRNA inhibits skeletal muscle aging by regulating miR-520d-5p

doi: 10.18632/aging.205279

Figure Lengend Snippet: The primer sequences for mRNAs, microRNAs and long non-coding RNAs (lncRNAs).

Article Snippet: Then, protein was transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, USA), blocked with 5% skim milk and incubated with primary antibodies of anti-Mef2c (Cal.

Techniques: Sequencing