Journal: Cell Reports Medicine

Article Title: Vaccine-induced ICOS + CD38 + circulating Tfh are sensitive biosensors of age-related changes in inflammatory pathways

doi: 10.1016/j.xcrm.2021.100262

Figure Lengend Snippet:

Article Snippet: Anti-human Survivin (REA459) , Miltenyi , Cat # 130-106-741; RRID: AB_2653606.

Techniques: Recombinant, Staining, Gene Expression, Software

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Proscillaridin A Promotes Oxidative Stress and ER Stress, Inhibits STAT3 Activation, and Induces Apoptosis in A549 Lung Adenocarcinoma Cells

doi: 10.1155/2018/3853409

Figure Lengend Snippet: PSD-A induces mitochondrial dysfunctions and ER stress in A549 cells. (a) Cells were treated with indicated concentrations of PSD-A for 24 h. Cell lysates were prepared and subjected to Western blot for the expressions of Bax, Bcl-2, Bcl-xl, Xiap, and survivin. GAPDH was used as loading control. (b) Cells were treated with PSD-A for 24 h in the presence or absence of BAPTA-AM (10 μ M), and MMP was determined by staining the cells with JC-1 according to the product's instructions. Data are expressed as mean ± SEM ( n = 3). Columns not sharing the same superscript letters differ significantly ( P < 0.05). (c, d) Cells were exposed to the indicated concentrations of PSD-A for 24 h. Total cell lysates were extracted, and expressions of p-eIF2 α , eIF2 α , CHOP, ATF4, GRP90, and GRP78 were measured by Western blot analyses. (e) Cells were treated with PSD-A for 24 h, and activity of caspase-4 was measured according to the kit's instructions. PSD-A increased the activity of caspase-4. Data are expressed as mean ± SEM ( n = 3). Columns not sharing the same superscript letters differ significantly ( P < 0.05).

Article Snippet: The primary antibodies for cleaved caspases (3 and 9), cleaved PARP, p-STAT3 (Tyr705), STAT3, p-SRC, and SRC were obtained from Cell Signaling Technology (Beverly, MA) while primary antibodies for Bax, Bcl-2, Bcl-xl, Xiap, survivin, ATF4, eIF2 α , GRP78, GRP98, GAPDH, SHP-1, SHP-2, PTEN, and TrxR1 were obtained from Proteintech (Wuhan, China).

Techniques: Western Blot, Control, Staining, Activity Assay

Journal: PeerJ

Article Title: Immune-related hub genes in intrauterine adhesions: a bioinformatics approach

doi: 10.7717/peerj.20035

Figure Lengend Snippet: (A) Representative IHC images showing protein expression levels of IGF1, WNT5A, BIRC5, and GDF7 in human endometrial tissues from IUA patients versus healthy controls (scale bar: 100 µm). (B) Corresponding IHC results in a mouse IUA model. (C) Quantitative analysis of IHC staining intensity in human tissues. (D) Quantitative analysis of IHC staining intensity in mouse tissues. Statistical significance was determined by Mann–Whitney U test (* P < 0.05, ** P < 0.01).

Article Snippet: The primary antibodies included IGF1 (Proteintech, #28530-1-AP; human 1:300, mouse 1:800), WNT5A (Proteintech, #55184-1-AP; human 1:800, mouse 1:1200), BIRC5 (ABclonal, #A1551; human 1:800, mouse 1:50), and GDF7 (Abcam, #ab189928, Clone EPR16000 ; human/mouse 1:1500).

Techniques: Expressing, Immunohistochemistry, MANN-WHITNEY

Journal: Frontiers in Cell and Developmental Biology

Article Title: Disruption of LTBP4 Inhibition-Induced TGFβ1 Activation Promoted Cell Proliferation and Metastasis in Skin Melanoma by Inhibiting the Activation of the Hippo-YAP1 Signaling Pathway

doi: 10.3389/fcell.2021.673904

Figure Lengend Snippet: LTBP4 inhibited the activation of Hippo signaling pathway in vitro . After shRNA-LTBP4#1 or #2 was transfected into SK-MEL-1 and VMM5A cells (A and B) the expressions of MST1 and MOB1 and the phosphorylation level of MST1 and MOB1 were detected by western blotting assay, (C) the mRNA levels of CTGF, Cyr61, and Birc5 were detected by RT-PCR assay, (D) the protein levels of CTGF, Cyr61, and Birc5 were detected by western blotting assay. After LTBP4 expressing plasmids transfected A101 and A375 cells (E and F) the expressions of MST1 and MOB1 and the phosphorylation level of MST1 and MOB1 were detected by western blotting assay, (G) the mRNA levels of CTGF, Cyr61, and Birc5 were detected by RT-PCR assay, and (H) the protein levels of CTGF, Cyr61, and Birc5 were detected by western blotting assay. β-actin was used as a load control. Data are presented as the mean ± standard deviation. ** p < 0.01 vs CTRL/or NC group.

Article Snippet: Antibody against LTBP4 was obtained from Shanghai Yu Bo Biotech Co.,Ltd. (Shanghai, China); antibody against cleaved caspase-3/Ki67/E-cadherin/YAP1/MST1/CTGF/Cyr61//TGFβR2 and phosph-YAP1/MST1 antibody was obtained from Abcam (Cambridge, UK); antibody against MOB1 and phospho-MOB1 were obtained from Cell Signaling Technology, Inc. (United States); antibody against Birc5 and phosphor-Birc5 were obtained from CUSABIO engineering co. Ltd. (Wuhan, Hubei, China); antibody against β-actin or LaminB were obtained from Beyotime (China).

Techniques: Activation Assay, In Vitro, shRNA, Transfection, Phospho-proteomics, Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Disruption of LTBP4 Inhibition-Induced TGFβ1 Activation Promoted Cell Proliferation and Metastasis in Skin Melanoma by Inhibiting the Activation of the Hippo-YAP1 Signaling Pathway

doi: 10.3389/fcell.2021.673904

Figure Lengend Snippet: LTBP4 suppressed secretion of active TGFβ1 as well as active TGFβ1-stimulated YAP1 nuclear-cytoplasmic translocation and the Hippo signaling inhibition. (A) Western blot, RT-PCR and ELISA analysis of TGFβRII expression (A) and total and active TGFβ1 (B) in shRNA-LTBP4#1 transfected SK-MEL-1 cells and LTBP4 expressing plasmids transfected A101 cells. A101D cells transfected with pcDNA3.1-LTBP4 were treated with 5 ng/ml TGF-β1, then, western blot analysis of YAP phosphorylation (C ), cytoplasmic YAP1 expression and nuclear YAP1 expression (D) , the luciferase activity of 8xGTIIC-luciferase analysis of YAP1 transcriptional activity (E) , and RT-PCR analysis of CTGF, Cyr61, and Birc5 expression (F) . β-actin and LaminB were used as a load control. Data are presented as the mean ± standard deviation. ** p < 0.01 vs CTRL/or NC group.

Article Snippet: Antibody against LTBP4 was obtained from Shanghai Yu Bo Biotech Co.,Ltd. (Shanghai, China); antibody against cleaved caspase-3/Ki67/E-cadherin/YAP1/MST1/CTGF/Cyr61//TGFβR2 and phosph-YAP1/MST1 antibody was obtained from Abcam (Cambridge, UK); antibody against MOB1 and phospho-MOB1 were obtained from Cell Signaling Technology, Inc. (United States); antibody against Birc5 and phosphor-Birc5 were obtained from CUSABIO engineering co. Ltd. (Wuhan, Hubei, China); antibody against β-actin or LaminB were obtained from Beyotime (China).

Techniques: Translocation Assay, Inhibition, Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing, shRNA, Transfection, Phospho-proteomics, Luciferase, Activity Assay, Control, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Disruption of LTBP4 Inhibition-Induced TGFβ1 Activation Promoted Cell Proliferation and Metastasis in Skin Melanoma by Inhibiting the Activation of the Hippo-YAP1 Signaling Pathway

doi: 10.3389/fcell.2021.673904

Figure Lengend Snippet: The functions of overexpression of LTBP4 regulating cleaved caspase-3, Ki67, E-cadherin, CTGF, Cyr61, and Birc5 were reversed by YAP1 OE or MST1 OE in melanoma cells. After LTBP4 expressing plasmids and YAP1/or MST1 expressing plasmids were co-transfected into SK-MEL-1 and A375 cells (A and B) the protein levels of cleaved caspase-3, Ki67, and E-cadherin in SK-MEL-1 cells were detected by western blotting assay (C and D) the protein levels of cleaved caspase-3, Ki67, and E-cadherin in A375 cells were detected by western blotting assay (E and F) the protein and mRNA levels of CTGF, Cyr61, and Birc5 in SK-MEL-1 cells were detected by western blotting and RT-PCR assays, and (G and H) the protein and mRNA levels of CTGF, Cyr61, and Birc5 in A375 cells were detected by western blotting and RT-PCR assays. β-actin was used as a load control. Data are presented as the mean ± standard deviation. ** p < 0.01 vs CTRL group and ## p < 0.01 vs LTBP4-OE group.

Article Snippet: Antibody against LTBP4 was obtained from Shanghai Yu Bo Biotech Co.,Ltd. (Shanghai, China); antibody against cleaved caspase-3/Ki67/E-cadherin/YAP1/MST1/CTGF/Cyr61//TGFβR2 and phosph-YAP1/MST1 antibody was obtained from Abcam (Cambridge, UK); antibody against MOB1 and phospho-MOB1 were obtained from Cell Signaling Technology, Inc. (United States); antibody against Birc5 and phosphor-Birc5 were obtained from CUSABIO engineering co. Ltd. (Wuhan, Hubei, China); antibody against β-actin or LaminB were obtained from Beyotime (China).

Techniques: Over Expression, Expressing, Transfection, Western Blot, Reverse Transcription Polymerase Chain Reaction, Control, Standard Deviation

Journal: Oncology Letters

Article Title: Identification of potential circular RNA biomarkers in lung adenocarcinoma: A bioinformatics analysis and retrospective clinical study

doi: 10.3892/ol.2022.13264

Figure Lengend Snippet: Relative expression and survival analysis of the six hub genes in LAC using the GEPIA database. Expression of (A) KIF2C, (B) KIF18B, (C) cytoskeleton-associated protein 2-like, (D) polo-like kinase 1, (E) maternal embryonic leucine zipper kinase and (F) and baculoviral IAP repeat-containing 5 was measured. Red color represents tumor tissues, and gray color represents normal tissues. (G) The circRNA-miRNA-hubgene sub-network was constructed. Diamonds represent circRNAs, triangles represent miRNAs and ovals represent mRNAs. *P<0.05. LAC, lung adenocarcinoma; GEPIA, Gene Expression Profiling Interactive Analysis; KIF, kinesin family member; circRNA, circular RNA; miRNA, microRNA.

Article Snippet: The sections were then treated with the 1:100-diluted KIF2C antibody (cat. no. TA503320; OriGene Technologies, Inc.), KIF18B (cat. no. ab121798; Abcam), CKAP2L (cat. no. ab122617; Abcam), PLK1 antibody (cat. no. TA500383; OriGene Technologies, Inc.), MELK antibody (cat. no. ab129373; Abcam), BIRC5 antibody (cat. no. TA301427; OriGene Technologies, Inc.) at 4°C overnight.

Techniques: Expressing, Construct, Gene Expression