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Image Search Results
Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Article Title: Human telomerase reverse transcriptase is a novel target of Hippo-YAP pathway.
doi: 10.1096/fj.201902147R
Figure Lengend Snippet: FIGURE 1 YAP increases hTERT expression. A and B, MCF7 cells were transiently transfected with YAP-WT, YAP-5SA expression vectors or empty vector (Vec) as a control. Relative mRNA levels were examined by qPCR, and protein levels were detected by western blot. As a well-established YAP target gene, CTGF was used as a positive control. C and D, qPCR analysis of relative mRNA levels, and western blot analysis of protein levels in YAP-WT, YAP-5SA, or empty vector transfected HEK293A cells. E and F, WT and LATS1/2 KO HEK293A cells were transfected with control siRNA (siNC) or siRNAs against YAP (siYAP-1, siYAP-2) for 48 h. The mRNA levels were analyzed by qPCR, and protein levels were determined by western blot. Values represent mean ± SEM from three independent experiments. *P < .05, **P < .01, ***P < .001 by one-way ANOVA (A and C) or two-way ANOVA (E) with Fisher's LSD test. Relative hTERT protein levels were quantified with ImageJ software and normalized to β-actin, as indicated at the bottom of the blot
Article Snippet: Primary antibodies against hTERT (ab32020; abcam), YAP (sc-101199; Santa Cruz Biotechnology), phospho-YAP (Ser127) (4911S; Cell Signaling Technology), LATS1 (3477T; Cell Signaling Technology), LATS2 (5888T; Cell Signaling Technology), TEA domain family transcription factor 4 (TEAD4) (ab58310; abcam),
Techniques: Expressing, Transfection, Plasmid Preparation, Control, Western Blot, Positive Control, Software
Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Article Title: Human telomerase reverse transcriptase is a novel target of Hippo-YAP pathway.
doi: 10.1096/fj.201902147R
Figure Lengend Snippet: FIGURE 3 YAP transcriptionally activates the hTERT promoter by interacting with TEAD. A and B, YAP induces hTERT expression in a TEAD-dependent manner. hTERT mRNA and protein levels were determined in HEK293A cells transfected with empty vector or YAP expressing vectors as indicated. Relative hTERT protein levels were quantified with ImageJ software and normalized to β-actin, as indicated at the bottom of the blot. C, HEK293A cells were transiently transfected with YAP-WT, 5SA, S94A expression vectors or empty vector, along with luciferase reporter vector containing a region of hTERT promoter (−951 to +58 bp). Luciferase activity was detected at 48 h after transfection. D, Luciferase reporter assay in HEK293A cells transfected with control (pGL2-Basic), hTERT promoter fragment (phTERT wt), or hTERT promoter fragment carrying mutation in potential TEAD binding motif (phTERT-TEAD mut), along with YAP-5SA expression vector. E, ChIP-qPCR analysis of YAP and TEAD4 enrichment at different regions of hTERT promoter in LATS1/2 KO HEK293A cells. GAPDH promoter was used as a negative control, and promoter of YAP target gene CTGF was used as a positive control. Values represent mean ± SEM from at least three independent experiments. *P < .05, **P < .01, ***P < .001 by one-way ANOVA (A, C and D) or two-way ANOVA (E) with Fisher's LSD test
Article Snippet: Primary antibodies against hTERT (ab32020; abcam), YAP (sc-101199; Santa Cruz Biotechnology), phospho-YAP (Ser127) (4911S; Cell Signaling Technology), LATS1 (3477T; Cell Signaling Technology), LATS2 (5888T; Cell Signaling Technology), TEA domain family transcription factor 4 (TEAD4) (ab58310; abcam),
Techniques: Expressing, Transfection, Plasmid Preparation, Software, Luciferase, Activity Assay, Reporter Assay, Control, Mutagenesis, Binding Assay, ChIP-qPCR, Negative Control, Positive Control