Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Loss of SPTBN1 Suppresses Autophagy Via SETD7-mediated YAP Methylation in Hepatocellular Carcinoma Initiation and Development

doi: 10.1016/j.jcmgh.2021.10.012

Figure Lengend Snippet: Loss of SPTBN1 inhibited autophagy by activating YAP in HCC cells. A and B , QRT-PCR. Huh-7 ( A ) and PLC/PRF/5 ( B ) cells were transiently transfected with siRNA to SPTBN1 or/and YAP for 48 hours and then analyzed. YAP siRNA reversed the expression of autophagy-related genes BECN1 and ATG4B that were downregulated by SPTBN1 knockdown (n = 3; ∗ P < .05, ∗∗ P < .01 vs siCON, # P < .05, ## P < .01 vs siSPTBN1). C and D , Western blot. The decreased ratio of LC3BII/LC3B I protein was reversed by YAP siRNA in Huh-7 ( C ) and PLC/PRF/5 ( D ) cells. Significance of the mean value difference was determined using a Student t test (∗∗ P < .01 vs siCON, ## P < .01 vs siSPTBN1). E , Analysis of autophagy level by Western blot. The decreased ratio of protein LC3BII/ LC3B I was reversed by YAP inhibitor verteporfin in the Huh-7 ( upper ) and PLC/PRF/5 ( lower ) HCC cells. F , The statistical analysis of Western blot in Figure E (n = 3; ∗∗ P < .01 vs siCON, # P < .05, ## P < .01 vs siSPTBN1). G , Autophagy and autophagic flux detection by fluorescence microscopy after GFP-RFP-LC3 LV transfection and HBSS induction for 24 hours. The reduced autophagy spots in response to SPTBN1 knockdown were reversed by YAP siRNA in Huh-7 ( left ) and PLC/PRF/5 ( right ) cells. White bars represent 10 μm.

Article Snippet: Cells were also treated with the YAP inhibitor verteporfin (10 μM, APExBIO, Houston, TX) after SPTBN1 siRNA transfection.

Techniques: Quantitative RT-PCR, Transfection, Expressing, Knockdown, Western Blot, Fluorescence, Microscopy

Journal: Molecular Medicine Reports

Article Title: COL1A1 expression induced by overexpression of both a 15-amino acid peptide from the fibrinogen domain of tenascin-X and integrin α11 in LX-2 cells

doi: 10.3892/mmr.2022.12846

Figure Lengend Snippet: Induction of COL1A1 expression by overexpression of both hTNX-FGFFFF and ITGA11 in addition to inhibitors. (A) Induction of COL1A1 expression by overexpression of both hTNX-FGFFFF and ITGA11 with a TGFBRI inhibitor (SB525334). DMSO (lanes 1, 2 and 3) and SB525334 (lane 3) were added to the culture medium (DMEM/0.5% FBS) after the transfection of expression plasmids for both hTNX-FGFFFF and ITGA11 in LX-2 cells (lanes 2 and 3). (B) Induction of COL1A1 expression by overexpression of both hTNX-FGFFFF and ITGA11 with a YAP inhibitor (verteporfin). DMSO (lanes 1, 2 and 3) and vertepofin (lane 3) were added to the culture medium (DMEM/0.5% FBS) after the transfection of expression plasmids for both hTNX-FGFFFF and ITGA11 in LX-2 cells (lanes 2 and 3). Subsequently, the cells were cultured followed by cell lysate extraction, RNA purification and reverse transcription-quantitative PCR. (A and B) The expression level of each gene [ ACTA2, COL1A1 and TGFB1 for (A) and ACTA2, COL1A1 and YAP1 for (B)] in the control (lane 1) was set to 1.0, and the relative expression level of each gene compared with that of the control (lane 1) is shown (n=3). Data are presented as the mean ± SD. *P<0.05, **P<0.01 vs. control (lane 1); ## P<0.01 vs. lane 2, one-way ANOVA with the Bonferroni post hoc test. COL1A1 , type I collagen α1 chain; TNX, tenascin-X; ITGA11, integrin α11; ACTA2 , α-smooth muscle actin; YAP, Yes-associated protein; hTNX-FGFFFF, GGLRIPFPRDCGEEM peptide from fibrinogen-related domain of human tenascin-X (hTNX-FG).

Article Snippet: To investigate the possible signaling pathway involved in the induction of COL1A1 expression by overexpression of both hTNX-FGFFFF and integrin α11 in LX-2 cells, a TGF-β receptor type 1 (TGFBRI) inhibitor (SB525334) (MedChemExpress) and a YAP inhibitor (verteporfin) (Cayman Chemical) were used.

Techniques: Expressing, Over Expression, Transfection, Cell Culture, Extraction, Purification, Reverse Transcription, Real-time Polymerase Chain Reaction, Control

Journal: Frontiers in Pharmacology

Article Title: Therapeutic effect of chinese herbal medicine gu-ben-hua-shi (AESS) formula on atopic dermatitis through regulation of yes-associated protein

doi: 10.3389/fphar.2022.929580

Figure Lengend Snippet: Inhibiting YAP expression could weaken the effect of AESS contained serum on AD-like cell model. (A) YAP, IκB-α, p-IκB-α, NF-κB p65 and p-NF-κB p65 expression. (B) Cell proliferation was examined by MTT. (C) Apoptosis was examined FACS analysis. (D) Adhesion to THP-1 cells (100 ×). (E) ICAM-1 and inflammatory cytokines (IFN-γ, IL-4, IL-17A, TGF-β and IL-10) mRNA expression. MTT for five replicates. Cell apoptosis, adhesion assay and qRT-PCR for three replicates. AD: AD-like cells were established by HaCaT cells culture with 10 ng/ml TNF-α/IFN-γ; AESS: AD-like cells culture with AESS contained serum; AESS + VP: AD-like cells culture with AESS contained serum and 2 μL YAP inhibitor verteporfin (VP). * p < 0.05, ** p < 0.01, *** p < 0.001 v. s. AD; # p < 0.05, ## p < 0.01, ### p < 0.001 v. s. AESS.

Article Snippet: The YAP inhibitor verteporfin (VP, 2 μl; Shanghai Yuanye Biotechnology Co., Ltd., China; ) was added to the cell culture medium to inhibit YAP expression in HaCaT cells.

Techniques: Expressing, Cell Adhesion Assay, Quantitative RT-PCR