Phospho-YAP (Ser127) Antibody - #AF3328

Fig. 4. The correlation between trans-differentiation and PIEZO1/YAP expression in MSCs. (A) Representative immunofluorescent images for the subcellular localization and activating states of YAP/TAZ in MSCs plating on GelMA and hard substrate (HS) before trans-differentiation. Scale bar:100 μm. (B) Representative immunofluorescent images for the activating states of YAP/TAZ and expression of S100β in GelMA and HS groups after trans-differentiation. Scale bar:100 μm. (C) Representative immunofluorescent images for the subcellular localization and activating states of YAP/TAZ and PIEZO1 in MSCs after trans-differentiation. Scale bar:10 μm. (D) Immunoblotted image for PIEZO2, PIEZO1, YAP/TAZ, p-YAP, GFAP, NGF, S100β in MSCs plating on GelMA and HS after co-culture with Ne–4C. (E) Densitometric analysis of blots showing the values of relevant proteins in GelMA and HS groups. (F) Immunoblotted image for PIEZO1, YAP/TAZ, p-YAP and GFAP in MSCs in co-culture system, after being treated with Yoda1 and GsMTx4. (G) Densitometric analysis of blots showing the values of relevant proteins in MSCs after being treated with Yoda1 and GsMTx4. (H) An indirect co-culture system including NE-4C in the upper chamber and MSCs plated on GelMA in the lower chambers.

Figure 4.| Upregulation of ST6GAL2 Rescues Tumorigenesis of FTC238 Cells and Resuppresses Hippo Signaling Pathway Activity(A–F) ST6GAL2 knockdown cells were transfected with ST6GAL2 overexpression vectors, and the proliferation,migration, and invasion capacities of FTC cells were enhanced. (G and H) Western blotting was performed to determine the levels of Hippo signaling molecules in FTC cells. *p < 0.05; scale bars, 20 mm.

Fig. 3Knockdown of HSP110 inhibits hypoxia-induced autophagy and YAP/TAZ-TEAD4 activity in mice. Relative mRNA level (a) and protein level (b, c) of HSP110 pulmonary arteries in lung tissues in each group (N = 8). d Double immunofluorescence staining of α-SMA (green) and HSP110 (red) in pulmonary arteries (N = 8). White scale bars, 50 μm; Yellow scale bars, 25 μm. White arrows pointed to α-SMA and HSP110 double-positive cells. e Protein levels of LC3II, LC3I, Beclin1, ATG5, ATG7 and p62 in pulmonary arteries (N = 8). f–h Quantitative analysis of relative protein ratio of LC3-II/I and relative protein level of Beclin1, p62, ATG5 and ATG7 (N = 8). i Double immunofluorescence staining of α-SMA (green) and Beclin 1 (red) in pulmonary arteries (N = 8). Scale bars, 50 μm. j Protein levels of p-YAP, YAP, p-TAZ and TAZ in pulmonary arteries (N = 8). k Quantitative analysis of relative protein ratio of p-YAP/t-YAP and p-TAZ/t-TAZ (N = 8). l–n Nuclear protein levels of YAP, TAZ and TEAD4 and quantitative analysis of relative protein level of nuclear YAP, TAZ and TEAD4 (N = 8). o Double immunofluorescence staining of α-SMA (green) and YAP (red) in pulmonary arteries (N = 8). White scale bars, 50 μm; Yellow scale bars, 25 μm. White arrows pointed to α-SMA and YAP double-positive cells. Data are means ± SD from 8 mice per group. *p 

FIGURE 6 Transfecting macrophages with a YAP siRNA further enhanced Ang II–induced macrophage M1 polarization and adhesion. (A) YAP siRNA efficiencies (n = 3, mean and S.D., t-test, &&P < 0.05 compared with siRNA NC;%%P < 0.05 compared with siRNA 860); (B,C) Western Blot of YAP after siRNA transfection *P < 0.05 compared with siRNA NC; (D) Flow cytometry analysis of CD68, CD86, and CD206 expression after 96 h co-culture in both groups. The data in the one-factor histogram represents the cells in the red circle in the upper scatter plot. HAECs + THP-1 + Ang II (1 μM, 24 h) + siRNA NC group: CD68, 41.94%; CD86, 45.73%; CD206, 9.72%. HAECs + THP-1 + Ang II + YAP siRNA group: CD68, 63.13%; CD86, 53.42%; CD206, 7.13%; (E–G) The proportion of CD68 +, CD86 +, and CD206 + cells, respectively (n = 3, mean and S.D., t-test, **P < 0.05 compared with the HAECs + THP-1 + siRNA NC group); (H) Fluorescence staining of adherent macrophages after YAP siRNA transfection. All cells were labeled with DAPI (blue), and the number of adherent macrophages was significantly decreased after transfection with the AT1R siRNA; (I) The ratio between the number of macrophages and the total number of cells (n = 3, mean and S.D., t-test, symbols are the same as in C,D,E).

Figure 6. The extent of apoptosis, glutathione (GSH) synthesis, and the expression of DNA damage-related proteins were assessed in cholangiocarcinoma cells (CCAs) subjected to YAP1 knockdown or YAP1 overexpression. Western blot was used to measure protein levels (n = 3). With cisplatin incubation, YAP1 knockdown and 1 mg/mL babaodan (BBD) treatment decreased the (a) Bcl-2 level and (b) increased bax level, while the change in (c) cle-caspase-3/caspase-3 levels with BBD treatment was not statistically significant. In the CCAs dealing with cisplatin, the expression levels of (d) p-YAP1/YAP1, (e) ATF4, and (f) SLC1A5 were decreased by YAP1 knockdown and BBD treatment. Additionally, the (g) γH2Ax level was increased and (h) the ERCC1 level was inhibited by YAP1 knockdown and BBD treatment. YAP1 overexpression antagonized the effect of BBD on these proteins. Representative protein bands are shown in (i), (j), and (k). (mean ± standard deviation) +p 

Fig. 5.| Influence of Sal-B on inflammatory response during YAP/TAZ/JNK signaling pathway in ECs. (A) Expression levels of YAP, TAZ, JNK, NF-κB and TNF-α were monitored by RT-PCR (n = 3). (B–C, F) Proteins (YAP, p-YAP, TAZ, p-TAZ, JNK, Nuclear NF-κB P65, Total P65 and TNF-α) in the pathway were detected by western blot (n = 3).