Journal: Journal of Gynecologic Oncology

Article Title: Downregulation of LINC01508 contributes to cisplatin resistance in ovarian cancer via the regulation of the Hippo-YAP pathway

doi: 10.3802/jgo.2021.32.e77

Figure Lengend Snippet: The cell function analysis are all chemically synthesized interference lncRNA and eukaryotic plasmid-mediated lncRNA transient transfection. These have limited gene silencing and overexpression efficiency and show that the correlation between LNC01508 and OC cisplatin resistance is not highly significant. Furthermore, the effect of eukaryotic plasmid-mediated lncRNA overexpression is better than the chemical synthesis of lncRNA. This suggests a stable transfection in vitro to provide knockout and overexpression effects. (A) qPCR detects a lower LINC01508 expression, and the CCK-8 assay measures the cell viability. IC 50 values in LINC01508 inhibition OV2008 cells. (B) qPCR inspects a higher LINC01508 expression, and the CCK-8 assay measures the cell viability. IC 50 values in LINC01508 overexpression C13K cells. (C and D) Colony formation assays were conducted to determine the cell proliferation ability for transfected OC cells. (E and F) GAPDH, Cyr61, ERCC1, and p-AKT protein levels were determined by Western blot. All experiments are repeated in triplicated, and data are shown as mean±SE. lncRNA, long non-coding RNAs; OC, ovarian cancer; qPCR, quantitative real-time polymerase chain reaction; CCK-8, cell counting kit-8; IC 50 , half maximal inhibitory concentration; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; Cyr61, cysteine-rich angiogenic inducer 61; ERCC1, excision repair cross complementing‐group 1; p-AKT, phospho-AKT; SE, standard error. * p<0.05; † p<0.01.

Article Snippet: The primary antibodies used were as follows: yes-associated protein (YAP) and cysteine-rich angiogenic inducer 61 (Cyr61) (sc-15407 and sc-13100, respectively; Santa Cruz Biotechnology, Dallas, TX, USA), GAPDH and phospho-AKT (p-AKT) (sc-166574 and sc-135650, respectively; Santa Cruz Biotechnology), phospho-YAP (p-YAP) and excision repair cross complementing‐group 1 (ERCC1) (4911 and 3885s, respectively; Cell Signaling Technology).

Techniques: Cell Function Assay, Synthesized, Plasmid Preparation, Transfection, Over Expression, Stable Transfection, In Vitro, Knock-Out, Expressing, CCK-8 Assay, Inhibition, Western Blot, Real-time Polymerase Chain Reaction, Cell Counting, Concentration Assay

Journal: Journal of Gynecologic Oncology

Article Title: Downregulation of LINC01508 contributes to cisplatin resistance in ovarian cancer via the regulation of the Hippo-YAP pathway

doi: 10.3802/jgo.2021.32.e77

Figure Lengend Snippet: (A) Western blot analyses of YAP and p-YAP protein expression in OC cells. (B and C) Cell survival rate and colony formation pictures of C13K cells after treatment of verteporfin and different doses of cisplatin. Data were shown as mean±SE of 3 experiments. (D) Cyr61 protein is marked by green fluorescence. Cyr61 protein was mainly localized in the cytoplasm. Nuclear staining with DAPI is shown in blue (magnification ×400). Representative images from 3 independent experiments are shown. OC, ovarian cancer; YAP, yes-associated protein 1; p-YAP, phospho-YAP; SE, standard error; Cyr61, cysteine-rich angiogenic inducer 61; DAPI, diamidino-2-phenylindole; DMSO, dimethyl sulfoxide. * p<0.05; † p<0.01, compared with DMSO group.

Article Snippet: The primary antibodies used were as follows: yes-associated protein (YAP) and cysteine-rich angiogenic inducer 61 (Cyr61) (sc-15407 and sc-13100, respectively; Santa Cruz Biotechnology, Dallas, TX, USA), GAPDH and phospho-AKT (p-AKT) (sc-166574 and sc-135650, respectively; Santa Cruz Biotechnology), phospho-YAP (p-YAP) and excision repair cross complementing‐group 1 (ERCC1) (4911 and 3885s, respectively; Cell Signaling Technology).

Techniques: Western Blot, Expressing, Fluorescence, Staining

Journal: Molecules

Article Title: Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins

doi: 10.3390/molecules25194395

Figure Lengend Snippet: Significantly upregulated proteins in human fetal retinal pigment epithelial cells (RPE) cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with 1% sericin compared to cells cultured in DMEM without sericin, sorted by decreasing fold change (FC). FC was calculated as follows: FC = ((mean of protein expressions in RPE cultured in DMEM with 1% sericin)/(mean of protein expressions in RPE cultured in DMEM without sericin)). N = 3. FC: Fold change.

Article Snippet: The blots were probed with antibodies for cysteine-rich angiogenic inducer 61 protein (CYR61; 1:100; Santa Cruz sc-374129, Santa Cruz Biotechnology, Dallas, TX, USA), laminin subunit β2 (1:500; Santa Cruz sc-133241), superoxide dismutase 1 (SOD1; 1:200; Santa Cruz sc-17767), cellular retinaldehyde-binding protein (CRALBP; 1:1000; Abcam ab-154898, Abcam, Cambridge, UK), premelanosome protein (PMEL; 1:250; Santa Cruz sc-33590), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1000; Santa Cruz sc-47724).

Techniques: Cell Culture, Modification

Journal: Molecules

Article Title: Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins

doi: 10.3390/molecules25194395

Figure Lengend Snippet: The table shows average fold change (FC) in protein levels upon incubation of primary human fetal retinal pigment epithelial cells (RPE) in Dulbecco’s Modified Eagle’s Medium (DMEM) with 1% sericin for seven days in comparison to control (cultured in DMEM alone). FC was calculated as follows: FC = ((mean of protein expressions in RPE cultured in DMEM with 1% sericin)/(mean of protein expressions in RPE cultured in DMEM without sericin)). Data are averages from triplicates. CYR61 = Cysteine-rich angiogenic inducer 61 protein.

Article Snippet: The blots were probed with antibodies for cysteine-rich angiogenic inducer 61 protein (CYR61; 1:100; Santa Cruz sc-374129, Santa Cruz Biotechnology, Dallas, TX, USA), laminin subunit β2 (1:500; Santa Cruz sc-133241), superoxide dismutase 1 (SOD1; 1:200; Santa Cruz sc-17767), cellular retinaldehyde-binding protein (CRALBP; 1:1000; Abcam ab-154898, Abcam, Cambridge, UK), premelanosome protein (PMEL; 1:250; Santa Cruz sc-33590), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1000; Santa Cruz sc-47724).

Techniques: Incubation, Modification, Cell Culture, Immunofluorescence, Western Blot

Journal: Molecules

Article Title: Sericin-Induced Melanogenesis in Cultured Retinal Pigment Epithelial Cells Is Associated with Elevated Levels of Hydrogen Peroxide and Inflammatory Proteins

doi: 10.3390/molecules25194395

Figure Lengend Snippet: Whole-cell lysates of human retinal pigment epithelial cells (RPE) cultured for seven days in Dulbecco’s Modified Eagle’s Medium (DMEM) with or without 1% sericin were subjected to Western blot experiments. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as loading control. Fold change (FC) was calculated as follows: FC = ((mean of protein expressions in RPE cultured in DMEM with 1% sericin)/(mean of protein expressions in RPE cultured in DMEM without sericin)). ( A ) Cysteine-rich angiogenic inducer 61 (CYR61); FC: 1.05; p -value: 0.90. ( B ); Laminin subunit β-2; FC: 1.11; p -value: 0.43. ( C ) Superoxide dismutase; FC: 1.36; p -value: 0.23. ( D ) Western blot. Error bars represent standard deviation of the mean. N = 3.

Article Snippet: The blots were probed with antibodies for cysteine-rich angiogenic inducer 61 protein (CYR61; 1:100; Santa Cruz sc-374129, Santa Cruz Biotechnology, Dallas, TX, USA), laminin subunit β2 (1:500; Santa Cruz sc-133241), superoxide dismutase 1 (SOD1; 1:200; Santa Cruz sc-17767), cellular retinaldehyde-binding protein (CRALBP; 1:1000; Abcam ab-154898, Abcam, Cambridge, UK), premelanosome protein (PMEL; 1:250; Santa Cruz sc-33590), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1000; Santa Cruz sc-47724).

Techniques: Cell Culture, Modification, Western Blot, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Upregulation of Yes-associated protein and transcriptional co-activator with PDZ-binding motif influences the behavior of LOVO human colon adenocarcinoma cells

doi: 10.3892/etm.2017.4962

Figure Lengend Snippet: YAP and/or TAZ overexpression changes the protein expression of tumor- or apoptosis-associated genes in LOVO cells. At 24 h after the transfection, CTGF, Cyr61, Bcl-2, Bax and caspase-3 protein expression levels were determined by using western blot analysis. Groups: NC, negative control; YAP mimics, cells transfected with YAP expression plasmids; TAZ mimics, cells transfected with TZA expression plasmids; YAP+TAZ, cells co-transfected with YAP and TAZ plasmids. YAP, Yes-associated protein; TAZ, transcriptional co-activator with PDZ-binding motif; CTGF, connective tissue growth factor; Cyr61, cysteine-rich angiogenic inducer 61; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein.

Article Snippet: Membranes were blocked at room temperature for 1 h in PBS solution supplemented with 0.1% Tween-20 and 5% fat-free powdered milk, and blotted with primary antibodies to connective tissue growth factor (CTGF; sc-101586; Santa Cruz Biotechnology, Inc.), cysteine-rich angiogenic inducer 61 (Cyr61; sc-374129; Santa Cruz Biotechnology, Inc.), B-cell lymphoma 2 (Bcl-2; AB112; Beyotime Institute of Biotechnology, Shanghai, China), Bcl-2-associated X protein (Bax; AB026; Beyotime Institute of Biotechnology), caspase-3 (AC030; Beyotime Institute of Biotechnology) (all used at 1:1,000 dilution) and GAPDH (AG019; Beyotime Institute of Biotechnology; 1:2,000 dilution) at 4°C overnight, followed by horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG antibody (A0192; Beyotime Institute of Biotechnology; 1:5,000 dilution) at room temperature for 2 h. Signals were detected by the enhanced chemiluminescence method (Thermo Fisher Scientific, Inc.).

Techniques: Over Expression, Expressing, Transfection, Western Blot, Negative Control, Binding Assay