Journal: Frontiers in Physiology

Article Title: Placental galectin-3 is reduced in early-onset preeclampsia

doi: 10.3389/fphys.2022.1037597

Figure Lengend Snippet: LGALS3 and LGALS3BP mRNA expression in first trimester placental stem cells differentiated into syncytiotrophoblast and extravillous trophoblasts. First trimester placental cytotrophoblast cells were differentiated into either syncytiotrophoblast or extravillous trophoblast (EVT) cells over 96 h. Syncytiotrophoblast differentiation was confirmed by increased expression of SDC1 (syncytiotrophoblast marker) (A) and decreased expression of CDH2 (cell border marker) (C) across time. LGALS3 (E) and LGALS3BP (G) mRNA expression with syncytiotrophoblast differentiation across 96 h. EVT differentiation was confirmed by increased expression of HLAG (EVT marker) (B) and reduced expression of TEAD4 (cytotrophoblast marker) (D) across time. LGALS3 (F) and LGALS3BP (H) mRNA expression with EVT differentiation over 96 h. All experiments were repeated n = 5 times in duplicate. Data is expressed as mean ± SEM; * p < 0.05, ** p < 0.01.

Article Snippet: RNA was converted to cDNA with high-capacity cDNA reverse transcriptase kit (Applied Biosystems, Life Technologies) as per manufacturer’s instructions using iCycler iQ5 machine (Biorad) with run conditions: 25°C for 10 min, 37°C for 60 min and 85°C for 5 min. Quantitative reverse transcriptase polymerase chain reaction (RT-PCR) measured the mRNA expression of genes; LGALS3 (Assay ID: Hs00173587_m1), LGALS3BP (Assay ID: Hs00174774_m1), TEAD4 (TEA Domain Transcription Factor 4, Assay ID: Hs01125032_m1), SDC1 (Syndecan 1, Assay ID: Hs00896423_m1) and HLAG (Human Leukocyte Antigen G, Assay ID: Hs03045108_m1) using Fluorescein amidite (FAM) labelled Taqman gene expression assays (Life Technologies) on the CFX 384 (Biorad, Hercules, CA) with Taqman fast advanced universal PCR mastermix (Applied Biosystems).

Techniques: Expressing, Marker

Journal: Progress in Orthodontics

Article Title: KAT6A/YAP/TEAD4 pathway modulates osteoclastogenesis by regulating the RANKL/OPG ratio on the compression side during orthodontic tooth movement

doi: 10.1186/s40510-024-00530-6

Figure Lengend Snippet: Mechanical force increased the expression of YAP and TEAD4. ( A ) Representative immunohistochemistry images of YAP and TEAD4 in compression side of PDL. The number of YAP + and TEAD4 + cells was increased on the compression side of PDL ( n = 3). The black arrow shows the direction of mechanical force. AB means the alveolar bone; PDL means the periodontal ligament; R means the root. *** P < 0.001 vs. control. Scale bar: 200 μm. ( B ) The western blot of YAP and TEAD4 after mechanical force application for 0 h, 6 h, 12 h, and 24 h. ( C ) The protein and mRNA levels of YAP and TEAD4 after mechanical force application in vitro ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. 0 h

Article Snippet: To inhibit the interaction between YAP and TEAD4, TED-347 (5 μM, HY-125,269, MedChemExpress, USA) was added to the medium.

Techniques: Expressing, Immunohistochemistry, Control, Western Blot, In Vitro

Journal: Progress in Orthodontics

Article Title: KAT6A/YAP/TEAD4 pathway modulates osteoclastogenesis by regulating the RANKL/OPG ratio on the compression side during orthodontic tooth movement

doi: 10.1186/s40510-024-00530-6

Figure Lengend Snippet: Mechanical force increased YAP association with TEAD4. ( A ) qPCR and western blot of TEAD4 after siYAP or YAP lentivirus treatment in PDLSCs. The protein and mRNA levels of TEAD4 were decreased and increased after siYAP or YAP lentivirus treatment, respectively ( n = 3). * P < 0.05 vs. control. ( B ) CO-IP was performed to detect YAP binding with TEAD4. ( C ) Representative immunofluorescence images of TEAD4 (red) and YAP (green) in PDLSCs. The nuc/cyto of YAP and the fluorescence colocalization area of YAP and TEAD4 were increased after mechanical force loading ( n = 3). Scale bar: 50 μm. ** P < 0.01, *** P < 0.001 vs. control. ( D ) Western blot of TEAD4 in PDLSCs. The protein level of TEAD4 was decreased after siKAT6A treatment ( n = 3). * P < 0.05 vs. siNC. ( E ) and ( F ) CO-IP was performed to detect YAP binding with TEAD4 and pan-acetylation after siKAT6A or WM-1119 treatment (24 h, 25 µM). The protein band indicated that the knockdown or inhibition of KAT6A decreased YAP association with TEAD4 and the pan-acetylation ( n = 3)

Article Snippet: To inhibit the interaction between YAP and TEAD4, TED-347 (5 μM, HY-125,269, MedChemExpress, USA) was added to the medium.

Techniques: Western Blot, Control, Co-Immunoprecipitation Assay, Binding Assay, Immunofluorescence, Fluorescence, Knockdown, Inhibition

Journal: Progress in Orthodontics

Article Title: KAT6A/YAP/TEAD4 pathway modulates osteoclastogenesis by regulating the RANKL/OPG ratio on the compression side during orthodontic tooth movement

doi: 10.1186/s40510-024-00530-6

Figure Lengend Snippet: The YAP/TEAD4 axis modulated osteoclastogenesis by regulating the RANKL/OPG ratio under mechanical force. ( A ) Representative Masson and H&E staining of the compression side of PDL. The black arrow shows the direction of mechanical force. The white dotted line indicates the shape of PDL. AB means the alveolar bone; PDL means the periodontal ligament; R means the root. Scale bar: 200 μm. ( B ) and ( C ) qPCR and western blot of RANKL and OPG after siYAP and siTEAD4 treatment in PDLCSs. The protein and mRNA levels of the RANKL/OPG ratio were decreased after siYAP or siTEAD4 treatment ( n = 3). * P < 0.05, ** P < 0.01 *** P < 0.01vs. siNC. D . Western blot and the quantitative analysis of the RANKL/OPG ratio in PDLSCs. The protein level of the RANKL/OPG ratio was upregulated after mechanical force loading, which could partly be reversed by TED-347 (24 h, 5 µM, n = 3). * P < 0.05, ** P < 0.01 vs. the force group

Article Snippet: To inhibit the interaction between YAP and TEAD4, TED-347 (5 μM, HY-125,269, MedChemExpress, USA) was added to the medium.

Techniques: Staining, Western Blot

Journal: Progress in Orthodontics

Article Title: KAT6A/YAP/TEAD4 pathway modulates osteoclastogenesis by regulating the RANKL/OPG ratio on the compression side during orthodontic tooth movement

doi: 10.1186/s40510-024-00530-6

Figure Lengend Snippet: Schematic illustration of the KAT6A/YAP/TEAD4 pathway modulating osteoclastogenesis during OTM by regulating the RANKL/OPG ratio

Article Snippet: To inhibit the interaction between YAP and TEAD4, TED-347 (5 μM, HY-125,269, MedChemExpress, USA) was added to the medium.

Techniques:

Journal: Nature neuroscience

Article Title: Epigenetic regulation of brain region-specific microglia clearance activity

doi: 10.1038/s41593-018-0192-3

Figure Lengend Snippet: (a,b) Representative immunofluorescence images are shown (NeuN+ neurons: red; cCASP3+: green; DAPI: blue). Dotted circles: cCASP3+/NeuN+ cell. Scale: 10 μm. (a) Quantification of cCASP3+ cells per cm 2 from 4mo control (striatum: mean=0, SEM=0; cerebellum: mean=2.500, SEM=1.443) or PLX-treated mice (striatum: mean=2.500, SEM=1.443; cerebellum: mean=119.2, SEM=20.02); p<0.0001, F=67.40; 14 cerebellum/striatum sections from n=4 mice/group. (b) Quantification of cCASP3+ cells per cm 2 from 4/5mo control (striatum: mean=0, SEM=0; cerebellum: mean=2.500, SEM=1.443) or Axl −/− Mertk −/− mice (striatum: mean=0, SEM=0; cerebellum: mean=33.17, SEM=8.355); p=0.0003, F=14.62;12 cerebellum/striatum sections from n=4 mice/group. Bar graphs with individual data points show mean ± SEM, one-way ANOVA with Tukey’s Multiple Comparison. (c) Horizontal bar graph shows relative expression (qPCR) of selected cbMg-enriched (orange) and stMg-enriched (purple) genes in microglia after 12 hours of exposure to vehicle or early apoptotic cells. Pparg (p=0.002, t 4 =7.144), Jdp2 (p<0.0001, t 4 =15.16), Rarg (p=0.001, t 4 =10.16), Tfec (p<0.0001, t 4 =11.47); Ahr (p=0.004, t 4 =6.033), En2 (p=0.025, t 4 =3.493), Tead4 (p=0.002, t 4 =7.013), Anxa2 (p<0.0001, t 4 =36.96), Colec12 (p=0.002, t 4 =7.498), Lilrb4 (p<0.0001, t 4 =14.04), Apoe (p=0.002, t 4 =6.964), Cd74 (p=0.001, t 4 =8.070), Ptch1 (p=0.037, t 4 =3.066), Clec7a (p=0.015, t 4 =4.064), Msr1 (p=0.011, t 4 =4.448), Lyz2 (p=0.024, t 4 =3.529), Ptplad2 (p=0.009, t 4 =4.809), Kdm6b (p<0.0001, t 4 =15.33), Kdm6a (p=0.004, t 4 =5.871), Hhex (p=0.025, t 4 =3.501), Esr1 (p=0.018, t 4 =3.885), Irf8 (p=0.001, t 4 =9.580), Sall1 (p=0.139, t 4 =1.842), Sall3 (p=0.449, t 4 =0.8381), Slc2a5 (p<0.0001, t 4 =15.30), Asb2 (p<0.0001, t 4 =25.35), Tmem119 (p=0.003, t 4 =6.721), Fscn1 (p<0.0001, t 4 =18.93), P2ry12 (p=0.733, t 4 =0.3659), and Fcrls (p=0.001, t 4 =10.31). Bar graphs show mean ± SEM, two-tailed unpaired t-test, n=3 wells of primary microglia cultures obtained from four 3mo mice. Experiment was independently reproduced 4 times.

Article Snippet: Probes used are Clec7a (Mm01183349_m1), Anxa2 (Mm01150673_m1), Lilrb4 (Mm01614371_m1), Msr1 (Mm00446214_m1), Cd74 (Mm01262765_g1), Apoe (Mm00437573_m1), Ptch1 (Mm00436026_m1), Ptplad2 (Mm01267670_m1), Lyz2 (Mm01612741_m1), Colec12 (Mm01236242_m1), Ahr (Mm00478932_m1), Jdp2 (Mm00473044_m1), Rarg (Mm00441091_m1), Pparg (Mm00440940_m1), Tfec (Mm01161234_m1), Tead4 (Mm01189836_m1), En2 (Mm00438710_m1), Asb2 (Mm01329892_m1), Irf8 (Mm00492567_m1), Hhex (Mm00433954_m1), Esr1 (Mm00433149_m1), Sall1 (Mm00491266_m1), Sall3 (Mm01265835_m1), Slc2a5 (Mm00600311_m1), Fscn1 (Mm00456046_m1), Fcrls (Mm01219431_m1), Tmem119 (Mm00525305_m1), P2ry12 (Mm01950543_s1), Ifnb1 (Mm00439552_s1), Mx1 (Mm00487796_m1), Ccl2 (Mm00441242_m1), Cxcl10 (Mm00445235_m1), Tnf (Mm00443258_m1), Kdm6b (Mm01332680_m1), Kdm6a (Mm00801998_m1), Gapdh (Mm99999915_g1).

Techniques: Immunofluorescence, Control, Comparison, Expressing, Two Tailed Test

Journal: iScience

Article Title: N 6 -methyladenosine-modified VGLL1 promotes ovarian cancer metastasis through high-mobility group AT-hook 1/Wnt/β-catenin signaling

doi: 10.1016/j.isci.2024.109245

Figure Lengend Snippet:

Article Snippet: For the colonization staining of TEAD4 and VGLL1, cells were washed three times with PBS, treated with PBS containing 1% Triton X-100 and then stained with anti-TEAD4 (Novus Biologicals, NBP1-32765, 1:200) and anti-VGLL1 antibodies (Proteintech, 10124-2-AP, 1:200).

Techniques: Recombinant, Transfection, Reverse Transcription, Immunoprecipitation, MTT Assay, Luciferase, Reporter Assay, Labeling, SYBR Green Assay, shRNA, Sequencing, Plasmid Preparation, Software, Microscopy