Journal: Cardiovascular Diabetology

Article Title: Expression of fourteen novel obesity-related genes in zucker diabetic fatty rats

doi: 10.1186/1475-2840-11-48

Figure Lengend Snippet: Primer and probes

Article Snippet: MTCH2 , Mitochondrial carrier homolog 2 , Rn01013168_m1.

Techniques: Derivative Assay, Variant Assay

Journal: Cardiovascular Diabetology

Article Title: Expression of fourteen novel obesity-related genes in zucker diabetic fatty rats

doi: 10.1186/1475-2840-11-48

Figure Lengend Snippet: Expression of genes with functions so far apart from the regulation of energy homeostasis. a) Relative expression of TFAP2B. * p < 0.05 vs. ZL KF and ZL MF; † p < 0.05 vs. ZDF KF and ZDF MF; # p < 0.05 vs. ZDF KF. b) Relative expression of ETV5. * p < 0.05 vs. ZL KF, ZL SF and ZL MF; † p < 0.05 vs. ZL HT and ZL MF; # p < 0.05 vs. ZDF KF and ZDF SF. c) Relative expression of MTCH2. * p < 0.05 vs. ZL KF, ZL MF and ZL HT; † p < 0.05 vs. ZL KF and ZL HT; # p < 0.05 vs. ZDF KF, ZDF SF and ZDF HT; ¶ p < 0.05 vs. ZDF SF; ‡ p < 0.05 vs. ZDF MF.

Article Snippet: MTCH2 , Mitochondrial carrier homolog 2 , Rn01013168_m1.

Techniques: Expressing

Journal: PLoS Genetics

Article Title: A Precisely Regulated Gene Expression Cassette Potently Modulates Metastasis and Survival in Multiple Solid Cancers

doi: 10.1371/journal.pgen.1000129

Figure Lengend Snippet: A) Real-time PCR quantification of siRNA mediated knockdown efficiency of five PGC genes ( p53CSV , MAP3K11 , MTCH2 , CPSF6 and SKIP ). The y-axis represents the percentage of relative silencing achieved by the different siRNA treatments. Relative silencing was calculated by comparing PGC gene expression levels between cells treated with either control or PGC target siRNAs. For each siRNA treatment, the expression levels of the PGC genes were normalized against the GADPH expression level. B) Representative photographs of AGS cells in the matrigel invasion assay. The left panel depicts control siRNA treated cells, while the right panel indicates p53CSV siRNA treated cells. Note the increased number of invading cells in the right panel. C) Summary graph of invasion effects caused by PGC gene silencing. Significant enhancements in cellular invasion were observed for p53CSV , MAP3K11 , MTCH2 , CPSF6 (* symbols, P<0.01). P-values were calculated using a one-tailed t-test. D) Summary graph of cell proliferation effects caused by PGC gene silencing. Significant reductions in cell proliferation were only observed for the SKIP siRNA treatments. P-values were calculated using a one-tailed t-test.

Article Snippet: Total RNA was reverse transcribed using Taqman Reverse Transcription Reagent kit (Applied Biosystems, Foster City, CA) and quantitative PCR was performed using the following Taqman probes: p53CSV (Hs00429934_g1); MAP3K11 (Hs00176759_m1); MTCH2 (Hs00819318_g1); CPSF6 (Hs00199668_m1); SKIP (Hs00273351_m1), on a 7900HT Fast Real time system (Applied Biosystems, Foster City, CA).

Techniques: Real-time Polymerase Chain Reaction, Knockdown, Gene Expression, Control, Expressing, Invasion Assay, One-tailed Test

Journal: Advanced Science

Article Title: MTCH2 Suppresses Thermogenesis by Regulating Autophagy in Adipose Tissue

doi: 10.1002/advs.202416598

Figure Lengend Snippet: Identification of MTCH2 as a conserved regulator of energy homeostasis. (A) A scheme of functional RNAi knockdown screen of 29 genes associated with weight loss using Drosophila melanogaster . (B) Z‐scores of TAG levels for each UAS‐RNAi line were presented. Z≥ 1.96 or ≤−1.96 (red dotted lines indicate threshold) was regarded as significant hits. (C) TAG levels were decreased in Mtch mutant flies fed normal, high sucrose or high‐fat diet ( n = 5 biological sample, 6 flies per sample). (D) Mtch2 mRNA expression was elevated in mice fed a high‐fat diet (DIO mice) compared to those mice fed a chow (lean mice) ( n = 6). (E,F) MTCH2 mRNA expression was lower in the abdominal scWAT of individuals with obesity than those without obesity ( n = 770 biological samples for E, n = 56 for (F). (G–J) A positive correlation existed between MTCH2 mRNA expression and body fat (G), BMI (H), waist‐to‐hip ratio (WHR) (I), and HOMA‐IR (J) in abdominal scWAT of individuals ( n = 49 biological samples for G, n = 770 biological samples for (H–J). (K) MTCH2 mRNA expression was significantly reduced in the abdominal scWAT of patients with obese after dietary intervention ( n = 46 biological samples). Data are represented as mean ± SEM. Two‐tailed unpaired Student's t ‐test (C–F), The Spearman's rank‐order correlation coefficient (r) (G–J), and two‐tailed paired t ‐test (K) were used. * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001.

Article Snippet: Heterozygous Mtch2 flox/+ mice were generated using CRISPR/Cas9 technology and obtained from Cyagen Biological Technology Co., Ltd. (Suzhou, China) on a C57BL/6J background.

Techniques: Functional Assay, Knockdown, Mutagenesis, Expressing, Two Tailed Test

Journal: Advanced Science

Article Title: MTCH2 Suppresses Thermogenesis by Regulating Autophagy in Adipose Tissue

doi: 10.1002/advs.202416598

Figure Lengend Snippet: Mtch2 AKO reduces fat depots and hepatic accumulation under HFD. (A) Mtch2 AKO did not affect body weight in mice fed chow ( n = 8–9). (B,C) Body weight was gained less in Mtch2 AKO mice under HFD feeding for 19 weeks. Representative image (B) and body weight growth curve ( n = 10, C). (D,E) Mtch2 AKO mice displayed a lower fat composition. Representative NMR analysis image (D) and fat mass composition by NMR (E). (F,G) Mtch2 AKO mice exhibited fewer fat depots. The dissected weight of total WAT, rWAT, scWAT and eWAT (F, n = 10), H&E staining and adipocyte size analysis of scWAT (G). (H,I) BAT was reduced in Mtch2 AKO mice. Representative image of dissected BAT (H, n = 10), the analyses of H&E staining and TEM on BAT and adipocyte size of BAT in H&E staining (I). (J–L) Fat accumulation was decreased in Mtch2 AKO mice. Representative image of the dissected liver (J, n = 10), hepatic TAG levels (K, n = 5), and liver H&E staining (L). (M) Mtch2 AKO mice exhibited enhanced insulin sensitivity in mice fed HFD for 18 weeks. Insulin tolerance test and quantified in the area under curves ( n = 8–10). Data are represented as mean ± SEM. Two‐way ANOVA followed by Bonferroni's multiple comparisons test (C,M), Two‐tailed unpaired t ‐test was used (E–K,M), * p < 0.05, ** p < 0.01, and *** p < 0.001, ns, not significant.

Article Snippet: Heterozygous Mtch2 flox/+ mice were generated using CRISPR/Cas9 technology and obtained from Cyagen Biological Technology Co., Ltd. (Suzhou, China) on a C57BL/6J background.

Techniques: Staining, Two Tailed Test

Journal: Advanced Science

Article Title: MTCH2 Suppresses Thermogenesis by Regulating Autophagy in Adipose Tissue

doi: 10.1002/advs.202416598

Figure Lengend Snippet: Mtch2 AKO increases energy expenditure by promoting BAT thermogenesis and scWAT browning in HFD‐fed mice. (A,B) Caloric intake was not altered in Mtch2 AKO mice. Daily food intake (A, n = 10) and accumulative food intake (B, n = 6). (C,D) Energy expenditure was enhanced in Mtch2 AKO mice ( n = 6). (E,F) Mtch2 AKO mice displayed higher surface temperature and core temperature under cold exposure (4 °C) at the age of 8 weeks. Representative infra‐red thermal image of lumbar back and BAT and qualification of temperatures of them ( n = 7 for E and 4 for F). (G,H) UCP1 protein levels were elevated in BAT and scWAT of Mtch2 AKO mice as shown in western blot (G, n = 4) and IHC staining of UCP1 (H). Data are represented as mean ± SEM. Two‐tailed unpaired t ‐test (A,C,E–G) or ANCOVA using body weight as covariate (D) was used. * p < 0.05, ** p < 0.01, and *** p < 0.001, ns, not significant.

Article Snippet: Heterozygous Mtch2 flox/+ mice were generated using CRISPR/Cas9 technology and obtained from Cyagen Biological Technology Co., Ltd. (Suzhou, China) on a C57BL/6J background.

Techniques: Western Blot, Immunohistochemistry, Two Tailed Test

Journal: Advanced Science

Article Title: MTCH2 Suppresses Thermogenesis by Regulating Autophagy in Adipose Tissue

doi: 10.1002/advs.202416598

Figure Lengend Snippet: Mtch2 AKO increases mitochondrial biogenesis and oxidative phosphorylation in adipose tissues of HFD‐fed mice. (A,B) The copies of mtDNA were increased in BAT (A) and scWAT (B) of Mtch2 AKO mice by qPCR qualification ( n = 5). (C) The number and structure of mitochondria were increased in BAT of Mtch2 AKO by TEM analysis. (D,E) Mitochondrial biogenesis was boosted in BAT (D, n = 5) and scWAT (E, n = 4–5) of Mtch2 AKO mice by qPCR analysis of typic markers. (F,G) Oxidative phosphorylation was enhanced in BAT (F, n = 5) and scWAT (G, n = 4–5) of Mtch2 AKO mice by qPCR analysis of typic markers. Data are represented as mean ± SEM. Two‐tailed unpaired t ‐test was used (A,B,D–G). * p < 0.05, ** p < 0.01, and *** p < 0.001, ns, not significant.

Article Snippet: Heterozygous Mtch2 flox/+ mice were generated using CRISPR/Cas9 technology and obtained from Cyagen Biological Technology Co., Ltd. (Suzhou, China) on a C57BL/6J background.

Techniques: Phospho-proteomics, Two Tailed Test

Journal: Advanced Science

Article Title: MTCH2 Suppresses Thermogenesis by Regulating Autophagy in Adipose Tissue

doi: 10.1002/advs.202416598

Figure Lengend Snippet: Mtch2 deficiency promotes autophagy. (A) Volcano plot of differentially expressed proteins in Mtch mutant flies (fold change > 1 and p < 0.05). (B) Ingenuity pathway analysis (IPA) of up‐regulated proteins in Mtch mutant flies. (C) Volcano plot of differentially expressed genes in BAT of HFD‐fed Mtch2 AKO mice (fold change > 1 and p < 0.05). (D) Venn diagram of the up‐regulated proteins in Mtch mutant flies and the up‐regulated genes in BAT of Mtch2 AKO mice. (E) Functional enrichment analysis of the overlapped up‐regulated genes in flies and mice. (F) Venn diagram of the down‐regulated genes in BAT of Mtch2 AKO mice and proteins in Mtch mutant flies (Left). The overlapped proteins localizing to the mitochondria were displayed (Right).

Article Snippet: Heterozygous Mtch2 flox/+ mice were generated using CRISPR/Cas9 technology and obtained from Cyagen Biological Technology Co., Ltd. (Suzhou, China) on a C57BL/6J background.

Techniques: Mutagenesis, Functional Assay

Journal: Advanced Science

Article Title: MTCH2 Suppresses Thermogenesis by Regulating Autophagy in Adipose Tissue

doi: 10.1002/advs.202416598

Figure Lengend Snippet: Mtch2 AKO promotes autophagy and lipolysis in the adipose tissues in HFD‐fed mice. (A–D) Autophagy was induced in BAT (A,C) and scWAT (B,D) of Mtch2 AKO mice by qPCR and western blot analysis of classic marker ( n = 4–6). (E) The number of autophagosomes and autophagolysosomes was increased in BAT of Mtch2 AKO mice by TEM analysis. (F,G) Mtch2 knockout reduced Bcl‐2 protein levels in BAT (F) and scWAT (G) in 19 weeks of HFD‐fed mice ( n = 6–8). (H–K) Lipolysis was increased in BAT (H,J) and scWAT (I,K) of Mtch2 AKO mice by qPCR and western blot analysis of lipolytic gene and protein expression ( n = 4–5). Data are represented as mean ± SEM. Two‐tailed unpaired t ‐test was used. * p < 0.05, ** p < 0.01 and *** p < 0.001, ns, not significant.

Article Snippet: Heterozygous Mtch2 flox/+ mice were generated using CRISPR/Cas9 technology and obtained from Cyagen Biological Technology Co., Ltd. (Suzhou, China) on a C57BL/6J background.

Techniques: Western Blot, Marker, Knock-Out, Expressing, Two Tailed Test

Journal: Advanced Science

Article Title: MTCH2 Suppresses Thermogenesis by Regulating Autophagy in Adipose Tissue

doi: 10.1002/advs.202416598

Figure Lengend Snippet: Mtch2 AKO promotes thermogenesis through the Bcl‐2‐autophagy pathways. (A) Autophagy was increased in 3T3‐L1 preadipocytes following Mtch2 knockout by CRISPR ( n = 4). (B) TAG levels were reduced in 3T3‐L1 mature adipocytes following Mtch2 knockout by CRISPR‐Cas9 technique ( n = 3). (C) UCP1 protein levels were elevated in 3T3‐L1 preadipocytes and 3T3‐L1 mature adipocytes following Mtch2 knockout by CRISPR ( n = 4). (D) CQ treatment (50 µ m ) increased autophagy but reduced UCP1 protein levels in 3T3‐L1 mature adipocytes following Mtch2 knockout by CRISPR ( n = 4). (E) MG132 treatment (10 µ m ) increased Bcl‐2 protein levels but reduced UCP1 protein levels in 3T3‐L1 mature adipocytes following Mtch2 knockout by CRISPR ( n = 3). (F) A summary of MTCH2 suppresses thermogenesis via Bcl2‐mediated autophagy in adipose tissues. MTCH2 functions as a negative regulator of autophagy. MTCH2 knockout induces Bcl‐2 protein degradation to enhance autophagy, leading to increased lipolysis, elevated UCP1 protein levels, and promotion of BAT thermogenesis and scWAT browning (Created with BioGDP.com). Data are represented as mean ± SEM. Two‐tailed unpaired t ‐test was used. * p < 0.05, ** p < 0.01 and *** p < 0.001, ns, not significant.

Article Snippet: Heterozygous Mtch2 flox/+ mice were generated using CRISPR/Cas9 technology and obtained from Cyagen Biological Technology Co., Ltd. (Suzhou, China) on a C57BL/6J background.

Techniques: Knock-Out, CRISPR, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: BRAF-Mutated Melanoma Cell Lines Develop Distinct Molecular Signatures After Prolonged Exposure to AZ628 or Dabrafenib: Potential Benefits of the Antiretroviral Treatments Cabotegravir or Doravirine on BRAF-Inhibitor-Resistant Cells

doi: 10.3390/ijms252211939

Figure Lengend Snippet: Characterization of phenotypes in melanoma cell populations resistant to dabrafenib or AZ628. This figure illustrates various features of melanoma cell populations with respect to signaling pathways, cell cycle regulation, and susceptibility to cell death. Receptor tyrosine kinase (RTK): including tyrosine-protein kinase receptor UFO (AXL) and mesenchymal–epithelial transition factor (c-Met). RAS/RAF/MAPK signaling pathway: detailing expression levels of V-raf murine sarcoma viral oncogenes (RAF) including BRAF and CRAF isoforms, as well as total extracellular signal-regulated kinases (ERK) expression. Pro-proliferative proteins: including phosphorylation levels of retinoblastoma protein (pRb) and M-phase inducer phosphatase 3 (pCDC25C), along with cyclin D1 and cyclin A2 expression. Apoptosis markers: displaying levels of cleaved poly (ADP-ribose) polymerase (cPARP) and mitochondrial carrier homolog 2 (MTCH2). Ferroptosis markers: highlighting the expression of transferrin and heme oxygenase-1 (HO-1), both of which contribute to increased intracellular iron. Levels of ferritin heavy chain 1 (FTH1), glutathione peroxidase 4 (GPX4), and cystine-glutamate antiporter (SLC7A11) are also presented.

Article Snippet: Following blocking, the membranes were incubated overnight at 4 °C on a shaker in a 5% BSA solution containing the following primary antibodies: Transferrin (A1448), Ferritin heavy chain (A19544), cleaved PARP-P25 (A19612), CDKN1B/p27KIP1 (A19095), CDKN2A/p16INK4a (A11651), Cyclin A2 (A19036), Bcl-xL (A0209), Heme Oxygenase 1 (A19062), ERK 1/2 (A16686), pERK 1/2 (AP0974), SLC7A11/xCT (A2413), C-Raf (A19638) from ABclonal (Woburn, MA, USA); Cyclin D1 (GTX634347), MTCH2 (GTX130324), β-actin (GTX124214), p21 Cip (GTX29543), CDC25C phospho Ser216 (GTX128153), BAX (GTX109683), c-Met (GTX100637), B-Raf1 (GTX100913) from Genetex (Alton Parkway, Irvine, CA, USA); Bcl-2 (#15071), pRb (#8516) from Cell Signaling Technology (Danvers, MA, USA) and AXL (13196-1-AP), GPX4 (67763-1) from ProteinTech (Manchester, UK).

Techniques: Expressing