Journal: PLoS ONE

Article Title: In Vivo Analysis of MEF2 Transcription Factors in Synapse Regulation and Neuronal Survival

doi: 10.1371/journal.pone.0034863

Figure Lengend Snippet: ( A ) Detection of Mef2a and Mef2d transcripts by in situ hybridization in littermate CTL and Mef2a/d DKO (DKO) mice. Arrow indicates hippocampus. ( B ) Expression level of MEF2 transcription factors in the hippocampus of Mef2a/d DKO mice as detected by quantitative RT-PCR. RNA was isolated from the hippocampus of 4 week old animals (n=3/genotype), and then expression determined by quantitative PCR. The fold change in RNA was calculated using the comparative Ct method, normalizing to GAPDH as a control. ( C ) Mef2a/d DKO mice exhibit impaired motor coordination as assessed by falling off the accelerating rotarod faster than littermate CTLs (F 1,7 =27.64, P<0.05; CTL, n=8; Mef2a/d DKO , n=10), posthoc analysis demonstrated that trial numbers 3–8 were significantly different between the Mef2a/d DKO and CTL mice. ( D ) Mef2a/d DKO mice show normal associative learning, as assessed by context- and cue-dependent fear conditioning (CTL, n=8; Mef2a/d DKO , n=10). ( E ) Representative images of Golgi-stained CA1 pyramidal neurons and a bar graph comparing the number of dendritic spines per 10 µm length of dendrite from littermate CTL and Mef2a/d DKO mice (n=31 dendritic segments from 31 neurons of two CTL mice and n=68 dendritic segments from 68 neurons of four Mef2a/d DKO mice). ( F ) CA1 LTP induction and maintenance is normal in DKO mice (CTL, n=3; Mef2a/d DKO , n=3). ( G ) Input-output relations are normal in Mef2a/d DKO mice (CTL, n=4; Mef2a/d DKO , n=4; solid lines indicate lines of best fit.). ( H ) Paired-pulse facilitation is increased in Mef2a/d DKO mice at Inter-Stimulus Interval 20, 30, 50, 100, 200 and 400 (mS) compared to littermate CTLs, suggesting decreased presynaptic release probability (CTL, n=8; Mef2a/d DKO , n=7). All data is shown as mean ± SEM.

Article Snippet: To assess MEF2B, C & D, Taqman probes (ABI) were used: Mef2b, Mm00484953_g1; Mef2c, Mm01344728_m1; Mef2d, Mm00504931_m1; Gapdh, Mm99999915_g1.

Techniques: In Situ Hybridization, Expressing, Quantitative RT-PCR, Isolation, Real-time Polymerase Chain Reaction, Control, Staining

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: Molecular medicine reports

Article Title: Optimization and enrichment of induced cardiomyocytes derived from mouse fibroblasts by reprogramming with cardiac transcription factors.

doi: 10.3892/mmr.2017.8285

Figure Lengend Snippet: Figure 1. Candidate factors for cardiomyocyte induction. TTFs were transduced with individual lentiviruses expressing (A) GATA4 (B) MEF2C (C) TBX5 and (D) HAND2, and were then compared with untransduced TTFs, which were used as the control. Cells were immunostained with corresponding antibodies and DAPI. Scale bars, 100 µm. TTFs, tail‑tip fibroblasts; GATA4, GATA binding protein 4; MEF2C, myocyte‑specific enhancer factor 2C; TBX5, T‑box transcription factor 5; HAND2, heart‑ and neural crest derivatives‑expressed protein 2.

Article Snippet: The primary antibodies against GATA4 (1:100, cat. no. sc-25310), MEF2C (1:100, cat. no. sc-13268), TBX5 (1:100, cat. no. sc-376952), HAND2 (1:100, cat. no. sc-9409), α-myosin heavy chain (MHC, 1:100, cat. no. sc-20641) (all from Santa Cruz Biotechnology, Inc., Dallas, TX, USA), α-sarcomeric actinin (1:100, cat. no. BM0003; Boster Biological Technology, Wuhan, China) and cardiac troponin T (cTnT, 1:200, cat. no. MAB-0374; Fuzhou Maixin Biotechnology, Fuzhou, China) were applied without washing and incubated at 4 ̊C overnight.

Techniques: Transduction, Expressing, Control, Binding Assay