Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Proliferation of hepatic stellate cells, mediated by YAP and TAZ, contributes to liver repair and regeneration after liver ischemia-reperfusion injury.

doi: 10.1152/ajpgi.00153.2017

Figure Lengend Snippet: Fig. 5. Hippo pathway and Yes-associated protein/transcriptional coactivator with PDZ-binding motif (YAP/TAZ) target protein expression in postischemic liver after ischemia-reperfusion (I/R). A: I/R leads to reduced phosphorylation of the Hippo pathway proteins, large tumor suppressor 1 (LATS1) and Mps one binder 1 (MOB1). B: I/R leads to increased expression of the YAP/TAZ target genes, CTGF and survivin. Lysates of postischemic liver tissue were analyzed by Western blot. Results were quantitated by image analysis. Data are means SE with n 3. *P 0.05 compared with sham mice.

Article Snippet: The antibodies were as follows: PCNA (sc-56; Santa Cruz Biotechnology, Dallas, TX), desmin (ab15200; Abcam, Cambridge, MA), F4/80 (ab6640, Abcam), -smooth muscle actin ( -SMA) (ab5694, Abcam), YAP (no. 14074; Cell Signaling Technology, Danvers, MA), TAZ (HPA007415; Sigma-Aldrich, St. Louis, MO), CK19 (TROMA-III; DSHB, Iowa City, IO), and CK8/18 (ab 53280; Abcam).

Techniques: Binding Assay, Expressing, Phospho-proteomics, Western Blot

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Proliferation of hepatic stellate cells, mediated by YAP and TAZ, contributes to liver repair and regeneration after liver ischemia-reperfusion injury.

doi: 10.1152/ajpgi.00153.2017

Figure Lengend Snippet: Fig. 6. Yes-associated protein/transcriptional coactiva- tor with PDZ-binding motif (YAP/TAZ) and Hippo pathway regulation in nonischemic liver after isch- emia-reperfusion (I/R). A: liver histopathology (H and E staining) in nonischemic liver after I/R showed no changes in liver architecture and no expansion of non- parenchymal cells. Original magnification is 100. B: YAP and TAZ expression (immunohistochemistry) in nonischemic liver lobes after 96 h of reperfusion was minimal in hepatocytes. Original magnification is 200. PV, portal vein; CV, central vein. C: no change in activation of Hippo pathway during I/R injury. Lysates of the nonischemic liver tissue were analyzed by Western blot and stained with phosphorylated large tumor suppressor 1 (LATS) and phosphorylated Mps one binder 1 (MOB1). D: expression of the YAP/TAZ target genes, CTGF and survivin, were assessed in lysates of nonischemic liver by Western blot. Results were quantitated by image analysis. Data are means SE with n 3. *P 0.05 compared with sham mice.

Article Snippet: The antibodies were as follows: PCNA (sc-56; Santa Cruz Biotechnology, Dallas, TX), desmin (ab15200; Abcam, Cambridge, MA), F4/80 (ab6640, Abcam), -smooth muscle actin ( -SMA) (ab5694, Abcam), YAP (no. 14074; Cell Signaling Technology, Danvers, MA), TAZ (HPA007415; Sigma-Aldrich, St. Louis, MO), CK19 (TROMA-III; DSHB, Iowa City, IO), and CK8/18 (ab 53280; Abcam).

Techniques: Binding Assay, Histopathology, Staining, Expressing, Immunohistochemistry, Activation Assay, Western Blot

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Proliferation of hepatic stellate cells, mediated by YAP and TAZ, contributes to liver repair and regeneration after liver ischemia-reperfusion injury.

doi: 10.1152/ajpgi.00153.2017

Figure Lengend Snippet: Fig. 7. Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) contribute to hepatic stellate cell (HSC) proliferation after ischemia-reperfusion (I/R). Mice were injected intraperitoneally with 10% dimethylsulfoxide solution (control) or 100 mg/kg verteporfin after 24 h of reperfusion and every 24 h thereafter. After 96 h of reperfusion, postischemic liver was analyzed. A: verteporfin treatment decreased HSC proliferation in ischemic liver after I/R. HSC proliferation was determined by dual immunofluorescence staining for desmin (red) and proliferating cell nuclear antigen (PCNA, green). DAPI was used for nuclear staining. Representative HSCs positive for PCNA are marked by arrows. Quantitative analysis of PCNA labeling was calculated as a percentage of PCNA-positive HSCs. Original magnification is 630. B: verteporfin treatment decreased the number of HSCs in postischemic liver after I/R. Desmin-labeled cells were measured as a percentage of total area. Original magnification is 200. C: verteporfin treatment decreased hepatocyte proliferation in postischemic liver after I/R. Hepatocyte proliferation was determined by immunohistochemical staining for PCNA. Original magnification is 200. D: verteporfin treatment decreased growth factors in postischemic liver after I/R. Liver hepatocyte growth factor (HGF) and amphiregulin levels were analyzed by ELISA. E: verteporfin treatment decreased the expression of survivin and Ankrd1, but not connective tissue growth factor (CTGF). The lysates of ischemic liver were analyzed by Western blot. Results were quantitated by image analysis. Data are means SE with n 3–4. *P 0.05 compared with control group.

Article Snippet: The antibodies were as follows: PCNA (sc-56; Santa Cruz Biotechnology, Dallas, TX), desmin (ab15200; Abcam, Cambridge, MA), F4/80 (ab6640, Abcam), -smooth muscle actin ( -SMA) (ab5694, Abcam), YAP (no. 14074; Cell Signaling Technology, Danvers, MA), TAZ (HPA007415; Sigma-Aldrich, St. Louis, MO), CK19 (TROMA-III; DSHB, Iowa City, IO), and CK8/18 (ab 53280; Abcam).

Techniques: Binding Assay, Injection, Control, Staining, Labeling, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Proliferation of hepatic stellate cells, mediated by YAP and TAZ, contributes to liver repair and regeneration after liver ischemia-reperfusion injury.

doi: 10.1152/ajpgi.00153.2017

Figure Lengend Snippet: Fig. 8. Inhibition of Yes-associated protein/ transcriptional coactivator with PDZ-bind- ing motif (YAP/TAZ) by verteporfin de- creases hepatic stellate cell (HSC) prolifer- ation and target gene expression in vitro. HSCs were isolated from mouse liver and treated with verteporfin or control culture medium for 5 days. A: verteporfin treatment decreased HSC proliferation as measured by DNA incorporation (n 8). B: verteporfin treatment did not induce cytotoxicity (lac- tate dehydrogenase, LDH, release) of HSCs (n 6). C: effect of verteporfin treatment (5 nM) on the release of hepatocyte growth factor (HGF) and amphiregulin by HSCs (n 3). HGF and amphiregulin were mea- sured by ELISA. Open bars, control; solid bars, verteporfin. D: effect of verteporfin treatment (5 nM) on expression of the YAP/ TAZ target genes, CTGF, survivin, and Ankrd1, measured by Western blot. Results were quantitated by image analysis (n 3). *P 0.05 compared with control group.

Article Snippet: The antibodies were as follows: PCNA (sc-56; Santa Cruz Biotechnology, Dallas, TX), desmin (ab15200; Abcam, Cambridge, MA), F4/80 (ab6640, Abcam), -smooth muscle actin ( -SMA) (ab5694, Abcam), YAP (no. 14074; Cell Signaling Technology, Danvers, MA), TAZ (HPA007415; Sigma-Aldrich, St. Louis, MO), CK19 (TROMA-III; DSHB, Iowa City, IO), and CK8/18 (ab 53280; Abcam).

Techniques: Inhibition, Targeted Gene Expression, In Vitro, Isolation, Control, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

Journal: Cell reports

Article Title: Mutant RIT1 cooperates with YAP to drive an EMT-like lung cancer state

doi: 10.1016/j.celrep.2025.116185

Figure Lengend Snippet: (A) Growth in low attachment assay of isogenic SALE cells expressing RIT1 M90I and YAP1 8SA alone or in combination analyzed by CellTiterGlo. Data shown are the mean ± SD of 14–16 technical replicates. (B) Heatmap of gene expression data derived from bulk RNA-seq of isogenic SALE cells. Each row is a differentially expressed gene, and each column is a replicate of the cell line variant indicated. The top 100 up- and downregulated genes distinguishing combined RIT1 M90I /YAP1 8SA (RY) cells from YAP1 8SA cells were determined by marker selection based on the mean difference of RY replicates compared to YAP1 8SA replicates and then genes and samples were clustered using one minus Pearson correlation. (C) Volcano plot of differentially expressed genes from RNA-seq in SALE-RY cells versus parental SALE cells. p values shown were calculated by t tests of three biological replicates per cell line. Multiple hypothesis testing was performed using the false discovery rate (FDR) method. Red points: log 2 fold change (log 2 FC) > 1.5, false discovery rate (FDR) < 0.1; blue points: log 2 FC < −1.5, FDR < 0.1. (D) MSigDB overlap analysis of up- and downregulated genes from RNA-seq in RY cells compared to parental. The FDR is the false discovery rate analog of the hypergeometic p value after correction for multiple hypothesis testing. (E) Motif analysis of up- and downregulated genes from (C) using the MSigDB Transcription Factor Target (TFT) gene set. AP-1 and ZEB1 (AREB6) motifs are shown in bold. FDR was calculated as in (D). (F) Western blot of isogenic SALE cells using antibodies against RIT1 and AP-1 transcription factors cJUN and FOSL1/FRA1. Vinculin is used as a loading control. (G) Quantification of biological replicates ( n = 3) of cJUN abundance determined by western blot, normalized to loading control and parental cJUN abundance. Data shown are mean ± SEM. P, parental; R, RIT1 M90I ; Y, YAP1 8SA ; RY, RIT1 M90I/ YAP1 8SA . (H) Quantification of biological replicates ( n = 3) of FRA1 abundance determined by western blot, normalized to loading control and parental FRA1 abundance. Data shown are mean ± SEM. Labeling as in (G). See also and . ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by unpaired two-tailed t test.

Article Snippet: YAP1 (D8H1X) (WB) , Cell Signaling Technology , Cat #14710; RRID: AB_2798483.

Techniques: Expressing, Gene Expression, Derivative Assay, RNA Sequencing, Variant Assay, Marker, Selection, Western Blot, Control, Labeling, Two Tailed Test

Journal: Cell reports

Article Title: Mutant RIT1 cooperates with YAP to drive an EMT-like lung cancer state

doi: 10.1016/j.celrep.2025.116185

Figure Lengend Snippet: (A) Immunofluorescence and confocal images of SALE cells fixed and stained with anti-YAP and anti-cJUN antibodies and counterstained with DAPI. Scale bar, 100 μm. Inset, 50 μm. (B) Quantification of nuclear YAP fluorescence intensity determined by confocal microscopy. Data shown are mean ± SEM of four biological replicates. P, parental; R, RIT1 M90I ; Y, YAP1 8SA ; RY, RIT1 M90I /YAP1 8SA (C) Quantification of nuclear cJUN fluorescence intensity determined by confocal microscopy. Data shown are mean ± SEM of four biological replicates. Labeling as in (B). (D) Dual p-AP1 luciferase reporter assay in HEK293T cells transiently transfected with control vectors (V), RIT1 M90I (R), or YAP1 WT and YAP1 8SA in the presence or absence of RIT1 M90I . p-RL renilla luciferase was co-transfected and used for normalization. PMA was used as a positive control. Data shown are mean ± SEM of 3 technical replicates. Data are representative of at least three independent experiments. (E) Western blot of SALE-RY cells showing doxycycline-regulated induction of cJUN or a dominant-negative cJUN (TAM67). (F) In vivo xenograft tumor growth on doxycycline of the cells shown in (E). Data shown are mean ± SEM of 8–10 tumors per group. * P < 0.05 by one-way ANOVA. (G) Tumor weights of tumors shown in (F). Data shown are mean ± SEM. The cJUN wild-type group was not analyzed due to early euthanasia for tumor ulceration. ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 by unpaired two-tailed t test unless otherwise specified. See also .

Article Snippet: YAP1 (D8H1X) (WB) , Cell Signaling Technology , Cat #14710; RRID: AB_2798483.

Techniques: Immunofluorescence, Staining, Fluorescence, Confocal Microscopy, Labeling, Luciferase, Reporter Assay, Transfection, Control, Positive Control, Western Blot, Dominant Negative Mutation, In Vivo, Two Tailed Test

Journal: The American Journal of Pathology

Article Title: TEA Domain Transcription Factor 4 Is the Major Mediator of Yes-Associated Protein Oncogenic Activity in Mouse and Human Hepatoblastoma

doi: 10.1016/j.ajpath.2019.01.016

Figure Lengend Snippet: Primary Antibodies Used for IHC and WB

Article Snippet: Phosphorylated Yap , Cell Signaling Technology Inc. , 13619P , 1:1000 , WB.

Techniques: