Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: ADAMTS1 levels were increased in the serum of patients with myocardial fibrosis, in the hearts of mice after CFPMI and in cardiac fibroblasts stimulated with fibrotic factors. A . ADAMTS1 levels in clinical serum samples were evaluated via ELISA. N = 30. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. B . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. C . Survival rate of mice in each group. D . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. E and F . ADAMTS1 expression in mouse heart tissue was determined through Western blot and IHC. Human cardiac fibroblasts were treated with 10 ng/mL TGF-β1 or 0.1 μM Ang II for 12, 24, or 48 h to construct in vitro models. The cells were divided into four groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups. G and H . Western blot detection of the expression of ADAMTS1, Collagen I and FN. I. The expression levels of ADAMTS1, Collagen I and FN were measured via qRT‒PCR. After the optimal time point (48 h) was determined, the mice were further divided into the control, TGF-β1 and Ang II groups. J . ADAMTS1 expression in cells was assessed by IF. Human cardiac fibroblasts were treated with 10 ng/mL TGF-β1 or 0.1 μM Ang II for 15, 30, 60, 90, and 120 min. K. qRT-PCR detection of the mRNA expression of ADAMTS1, Collagen I and FN. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The blocked membranes were incubated overnight at 4 °C with the following primary antibodies: ADAMTS1 (ab236497, 1:2000, Abcam), Collagen I (ab270993, 1:1000, Abcam), FN ( AWA00143 , 1:1000, Abiowell), SMAD2 (12,570–1-AP, 1:5000, Proteintech), p-SMAD2 (ab280888, 1:1000, Abcam), α-SMA (ab5694, 1 μg/ml, Abcam), HDAC6 (128,341-AP, 1:1000, Proteintech), TGF-β1 ( AWA00083 , 1:1000, Abiowell), and GAPDH (10,494–1-AP, 1:5000, Proteintech) at 4 °C overnight.

Techniques: Enzyme-linked Immunosorbent Assay, Construct, Staining, Expressing, Western Blot, In Vitro, Control, Quantitative RT-PCR

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: SMAD2 regulated ADAMTS1 expression in human and mouse cardiac fibroblasts induced by TGF-β1. We used 10 ng/mL TGF-β1 to induce human and mouse cardiac fibroblasts for 15, 30, 60, 90, and 120 min. A and B . Western blot analysis of the expression of SMAD2, p-SMAD2 and ADAMTS1. Human and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 for 48 h, and the cells were divided into Control and TGF-β1 groups. C . Human and mouse cardiac fibroblasts were induced with 10 ng/mL TGF-β1 for 12 h, 24 h, 48 h, and 72 h. qRT‒PCR was performed to detect the mRNA levels of ADAMTS1, Collagen I, and FN. D . IF staining was conducted to determine the expression of p-SMAD2 and ADAMTS1 at 48 h. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. E . p-SMAD2 expression in mouse heart tissue was measured by IHC. F . ChIP verification of the interaction between SMAD2 and ADAMTS1. Then, we interfered with SMAD2 expression. The groups were the si-NC, si-SMAD2-1, and si-SMAD2-2 groups. G . SMAD2 expression was assessed via qRT‒PCR. After the best si-SMAD2 was selected, the cells were further divided into the si-NC, TGF-β1 (48 h), si-SMAD2, and TGF-β1 + si-SMAD2 groups. H . Western blot detection of ADAMTS1, Collagen I, and FN expression in cells. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The blocked membranes were incubated overnight at 4 °C with the following primary antibodies: ADAMTS1 (ab236497, 1:2000, Abcam), Collagen I (ab270993, 1:1000, Abcam), FN ( AWA00143 , 1:1000, Abiowell), SMAD2 (12,570–1-AP, 1:5000, Proteintech), p-SMAD2 (ab280888, 1:1000, Abcam), α-SMA (ab5694, 1 μg/ml, Abcam), HDAC6 (128,341-AP, 1:1000, Proteintech), TGF-β1 ( AWA00083 , 1:1000, Abiowell), and GAPDH (10,494–1-AP, 1:5000, Proteintech) at 4 °C overnight.

Techniques: Expressing, Western Blot, Control, Staining, Construct

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: Overexpression of ADAMTS1 enhanced the production of collagen fiber proteins in human and mouse cardiac fibroblasts induced by TGF-β1. We overexpressed ADAMTS1 and divided the cells into the oe-NC, TGF-β1 (48 h), oe-ADAMTS1, and TGF-β1 + oe-ADAMTS1 groups. A . Western blot detection of ADAMTS1, Collagen I, FN, and α-SMA expression. B-D . IF staining of Collagen I and FN expression. Additionally, we interfered with ADAMTS1; the cells were grouped as follows: si-NC, si-ADAMTS1-1, and si-ADAMTS1-2. E . ADAMTS1 expression was determined through qRT‒PCR. After the best si-ADAMTS1 was selected, the cells were further divided into the si-NC, TGF-β1 (48 h), si-ADAMTS1, and TGF-β1 + si-ADAMTS1 groups. F and G . Western blot detection of ADAMTS1, Collagen I, FN, and α-SMA expression. H and I . IF staining of Collagen I and FN expression. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The blocked membranes were incubated overnight at 4 °C with the following primary antibodies: ADAMTS1 (ab236497, 1:2000, Abcam), Collagen I (ab270993, 1:1000, Abcam), FN ( AWA00143 , 1:1000, Abiowell), SMAD2 (12,570–1-AP, 1:5000, Proteintech), p-SMAD2 (ab280888, 1:1000, Abcam), α-SMA (ab5694, 1 μg/ml, Abcam), HDAC6 (128,341-AP, 1:1000, Proteintech), TGF-β1 ( AWA00083 , 1:1000, Abiowell), and GAPDH (10,494–1-AP, 1:5000, Proteintech) at 4 °C overnight.

Techniques: Over Expression, Western Blot, Expressing, Staining

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: Knockdown of ADAMTS1 alleviated TGF-β1-induced fibrosis by downregulating HDAC6 protein expression. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. A . HDAC6 expression in mouse heart tissue was determined through IHC. After the best si-ADAMTS1 was selected, the cells were further divided into the si-NC and si-ADAMTS1 groups. B . ADAMTS1 and HDAC6 expression was assessed via qRT‒PCR. C . Western blot analysis of ADAMTS1 and HDAC6 expression. Furthermore, we overexpressed ADAMTS1 and divided the cells into the oe-NC, TGF-β1, oe-ADAMTS1, and TGF-β1 + oe-ADAMTS1 groups. D . Western blot detection of HDAC6 expression. After the si-ADAMTS1 group was selected, the cells were further divided into the si-NC, TGF-β1, si-ADAMTS1, and TGF-β1 + si-ADAMTS1 groups. E. Western blot detection of HDAC6 expression. Moreover, cells were treated with the HDAC6 inhibitor ACY1215 (5 μM) for 48 h and then divided into the si-NC, si-ADAMTS1, si-NC + ACY1215, and si-ADAMTS1 + ACY1215 groups. F. Western blot detection of TGF-β1, Collagen I, FN, and ADAMTS1 expression. Subsequently, experiments were performed in human and mouse cardiac fibroblasts with ADAMTS1 overexpression followed by treatment with the HDAC6 inhibitor ACY1215. G. Western blot analysis was conducted to detect the expression levels of ADAMTS1 and HDAC6. H. Western blot analysis was performed to determine the expression levels of TGF-β1, Collagen I, and FN. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The blocked membranes were incubated overnight at 4 °C with the following primary antibodies: ADAMTS1 (ab236497, 1:2000, Abcam), Collagen I (ab270993, 1:1000, Abcam), FN ( AWA00143 , 1:1000, Abiowell), SMAD2 (12,570–1-AP, 1:5000, Proteintech), p-SMAD2 (ab280888, 1:1000, Abcam), α-SMA (ab5694, 1 μg/ml, Abcam), HDAC6 (128,341-AP, 1:1000, Proteintech), TGF-β1 ( AWA00083 , 1:1000, Abiowell), and GAPDH (10,494–1-AP, 1:5000, Proteintech) at 4 °C overnight.

Techniques: Knockdown, Expressing, Construct, Western Blot, Over Expression

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: ADAMTS1 interacted with HDAC6 during fibrosis. A . Co-IP verification of the interaction of ADAMTS1 with HDAC6, with ADAMTS1 as the bait protein. In the experiment, we first captured the ADAMTS1 protein using a specific antibody and then used Co-IP technology to detect its interaction with HDAC6. B . Co-IP verification of the interaction of HDAC6 with ADAMTS1, with HDAC6 as the bait protein. In the experiment, we captured the HDAC6 protein using a specific antibody and then used Co-IP technology to detect its interaction with ADAMTS1. We used 10 ng/mL TGF-β1 to induce human and mouse cardiac fibroblasts for 48 h, and the cells were further divided into control and TGF-β1 groups. C . IF staining was performed to evaluate the colocalization of ADAMTS1 and HDAC6 in TGF-β1-treated human and mouse cardiac fibroblasts. D . Changes in the level of ubiquitinated HDAC6 protein in human and mouse cardiac fibroblasts transfected with oe-ADAMTS1/si-ADAMTS1 or oe-NC/si-NC in the presence of 10 μM MG132. E. In si-ADAMTS1-treated human and mouse cardiac fibroblasts, MG132 (10 μM) was added to detect the ubiquitination levels of ADAMTS1 protein. F. Human and mouse cardiac fibroblasts were treated with TGF-β1 and subjected to SMAD2 knockdown to determine the ubiquitination levels of ADAMTS1 protein. N = 3. *** P < 0.001

Article Snippet: The blocked membranes were incubated overnight at 4 °C with the following primary antibodies: ADAMTS1 (ab236497, 1:2000, Abcam), Collagen I (ab270993, 1:1000, Abcam), FN ( AWA00143 , 1:1000, Abiowell), SMAD2 (12,570–1-AP, 1:5000, Proteintech), p-SMAD2 (ab280888, 1:1000, Abcam), α-SMA (ab5694, 1 μg/ml, Abcam), HDAC6 (128,341-AP, 1:1000, Proteintech), TGF-β1 ( AWA00083 , 1:1000, Abiowell), and GAPDH (10,494–1-AP, 1:5000, Proteintech) at 4 °C overnight.

Techniques: Co-Immunoprecipitation Assay, Control, Staining, Transfection, Ubiquitin Proteomics, Knockdown

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: AAV-shRNA-ADAMTS1 treatment alleviated myocardial fibrosis and improved cardiac function after CFPMI. The mice were randomly divided into the Sham, CFPMI, and CFPMI + sh-ADAMTS1 groups. A . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. B . Survival rate of mice in each group. C . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. D . HDAC6 expression was determined through qRT‒PCR. E. Western blot analysis of the expression of TGF-β1, SMAD2, and p-SMAD2 in mouse heart tissue. F. Western blot detection of Collagen I, and FN expression. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The blocked membranes were incubated overnight at 4 °C with the following primary antibodies: ADAMTS1 (ab236497, 1:2000, Abcam), Collagen I (ab270993, 1:1000, Abcam), FN ( AWA00143 , 1:1000, Abiowell), SMAD2 (12,570–1-AP, 1:5000, Proteintech), p-SMAD2 (ab280888, 1:1000, Abcam), α-SMA (ab5694, 1 μg/ml, Abcam), HDAC6 (128,341-AP, 1:1000, Proteintech), TGF-β1 ( AWA00083 , 1:1000, Abiowell), and GAPDH (10,494–1-AP, 1:5000, Proteintech) at 4 °C overnight.

Techniques: shRNA, Staining, Expressing, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: AAV-shRNA-HDAC6 transfection combined with ADAMTS1 inhibitor treatment alleviated myocardial fibrosis and improved cardiac function after CFPMI. The mice were randomly divided into the Sham, CFPMI, CFPMI + anti-ADAMTS1, CFPMI + sh-HDAC6, and CFPMI + anti-ADAMTS1 + sh-HDAC6 groups. A . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. B . Survival rate of mice in each group. C and D . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. E. HDAC6 expression was determined through qRT‒PCR. F. Western blot analysis of the expression of TGF-β1, SMAD2, and p-SMAD2 in mouse heart tissue. G. Western blot detection of Collagen I and FN expression. N = 3. ** P < 0.01, *** P < 0.001

Article Snippet: The blocked membranes were incubated overnight at 4 °C with the following primary antibodies: ADAMTS1 (ab236497, 1:2000, Abcam), Collagen I (ab270993, 1:1000, Abcam), FN ( AWA00143 , 1:1000, Abiowell), SMAD2 (12,570–1-AP, 1:5000, Proteintech), p-SMAD2 (ab280888, 1:1000, Abcam), α-SMA (ab5694, 1 μg/ml, Abcam), HDAC6 (128,341-AP, 1:1000, Proteintech), TGF-β1 ( AWA00083 , 1:1000, Abiowell), and GAPDH (10,494–1-AP, 1:5000, Proteintech) at 4 °C overnight.

Techniques: shRNA, Transfection, Staining, Expressing, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: TGF-β1/SMAD2 regulated ADAMTS1 by mediating CFPMI through HDAC6 ubiquitination

Article Snippet: The blocked membranes were incubated overnight at 4 °C with the following primary antibodies: ADAMTS1 (ab236497, 1:2000, Abcam), Collagen I (ab270993, 1:1000, Abcam), FN ( AWA00143 , 1:1000, Abiowell), SMAD2 (12,570–1-AP, 1:5000, Proteintech), p-SMAD2 (ab280888, 1:1000, Abcam), α-SMA (ab5694, 1 μg/ml, Abcam), HDAC6 (128,341-AP, 1:1000, Proteintech), TGF-β1 ( AWA00083 , 1:1000, Abiowell), and GAPDH (10,494–1-AP, 1:5000, Proteintech) at 4 °C overnight.

Techniques: Ubiquitin Proteomics

Journal: Nature Communications

Article Title: Lactate derived from macrophages drives skin dermal fibroblasts phenotypic remodeling via MCT1-primed histone H3 lysine 23 lactylation in hypertrophic scar

doi: 10.1038/s41467-026-69388-y

Figure Lengend Snippet: A , B Extracellular (left panel) and intracellular (right panel) lactate levels in THP-1 cells, HUVECs, and NSFs cultured with varying substrate stiffness ( A ) or the treatment of TGF-β ( B ) ( n = 5 independent biological replicates). C , D ECAR of THP-1 cells with the treatment of varying substrate stiffness ( C ) or TGF-β ( D ) were monitored in real-time using the seahorse system. ECAR was used primarily to measure glycolysis (left panel) and assess cellular glycolysis, glycolytic capacity and glycolysis reserve (left and right panel, n = 5 independent biological replicates). E , F ECAR of HUVECs ( E ) and NSFs ( F ) with the treatment of different substrate stiffness were monitored in real-time using the seahorse system. ECAR was used primarily to measure glycolysis (left panel) and assess cellular glycolysis, glycolytic capacity and glycolysis reserve (left and right panel, n = 5 independent biological replicates). G Extracellular (left panel) and intracellular (right panel) levels in primary macrophages, vascular endothelial cells and dermal fibroblasts in NS and HS tissues ( n = 5 independent biological replicates). H ECAR of primary macrophages from NS and HS tissues were monitored in real-time using the seahorse system. ECAR was used primarily to measure glycolysis (left panel) and assess cellular glycolysis, glycolytic capacity and glycolysis reserve (left and right panel, n = 5 independent biological replicates). I , J Representative images of CD68 co-stained with HK2 ( I ) and LDHA ( J ) in normal skin and hypertrophic scar tissues (left panel); quantification of HK2 ( I ) and LDHA ( J ) intensity were shown in statistical diagram of CD68-positive cells (right panel). White arrowheads indicate the colocalization between HK2 or LDHA and CD68 in normal skin and hypertrophic scar tissues (scale bar = 100 and 20 μm, n = 50 cells per group). Data are presented as the mean ± SD. Two-tailed unpaired t-test was used for comparison in ( A – J ). Source data are provided as a Source Data file.

Article Snippet: The lactate production inhibitor Oxamate (10 mM) was used to pre-treat THP-1 cells and HUVECs prior to supernatant collection after 24 h. THP-1 cells and HUVECs were pretreated for 24 h with TGF-β (10 ng/ml, Proteintech, HZ-1011).

Techniques: Cell Culture, Staining, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: Lactate derived from macrophages drives skin dermal fibroblasts phenotypic remodeling via MCT1-primed histone H3 lysine 23 lactylation in hypertrophic scar

doi: 10.1038/s41467-026-69388-y

Figure Lengend Snippet: A Schematic showing the strategy for CM derived from different substrate stiffness induced THP-1 cells and HUVECs to stimulate NSFs. Created in BioRender. Rui, Z. ( https://BioRender.com/zfxf6v3 ). B , G Western blotting analysis of fibrotic markers of NSFs stimulated by 50 kPa substrate stiffness-induced THP-1 cells CM, followed by treatment with or without AZD3965 ( B ) or Oxamate ( G ) ( n = 3 independent biological replicates). C , H Representative immunofluorescence staining images of fibrotic markers of NSFs stimulated by 50 kPa substrate stiffness-induced THP-1 cells CM, followed by treatment with or without AZD3965 ( C ) or Oxamate ( H ) (scale bar = 50 μm, n = 3 independent biological replicates). D , I Representative scratch wound healing images at 0 h and 24 h of NSFs stimulated by 50 kPa substrate stiffness-induced THP-1 cells CM, followed by treatment with or without AZD3965 ( D ) or Oxamate ( I ) (scale bar = 100 μm, n = 5 independent biological replicates). E , J Statistical diagram of percentage of Ki-67-positive cells of NSFs stimulated by 50 kPa substrate stiffness-induced THP-1 cells CM, followed by treatment with or without AZD3965 ( E ) or Oxamate ( J ) ( n = 10 independent biological replicates). F , K RT–qPCR analysis of relative mRNA expression of profibrotic markers (Fibronectin, SPARC, POSTN, TNC, CD248 and SMAD2) of NSFs stimulated by 50 kPa substrate stiffness-induced THP-1 cells CM treated with AZD3965 ( F ) or Oxamate ( K ) ( n = 5 independent biological replicates). L Western blotting analysis of fibrotic markers of NSFs stimulated by TGF-β-induced THP-1 cells CM, followed by treatment with or without AZD3965 ( n = 3 independent biological replicates). M Representative immunofluorescence staining images of fibrotic markers (α-SMA, green; COL1A1, red) of NSFs stimulated by TGF-β-induced THP-1 cells CM, followed by treatment with or without AZD3965 (scale bar = 50 μm, n = 3 independent biological replicates). The samples derive from the same experiment and that blots were processed in parallel. Data are presented as the mean ± SD. One-way ANOVA analysis followed by two-tailed Tukey post hoc multi-comparison test was used for comparison in ( B , E , F , G , J , K and L ). Source data are provided as a Source Data file.

Article Snippet: The lactate production inhibitor Oxamate (10 mM) was used to pre-treat THP-1 cells and HUVECs prior to supernatant collection after 24 h. THP-1 cells and HUVECs were pretreated for 24 h with TGF-β (10 ng/ml, Proteintech, HZ-1011).

Techniques: Derivative Assay, Western Blot, Immunofluorescence, Staining, Quantitative RT-PCR, Expressing, Two Tailed Test, Comparison

Journal: Cancer Communications

Article Title: Ectopic CD11c Drives SMAD3-Mediated Aberrant Antigen Presentation and Epithelial–Mesenchymal Transition in Esophageal Squamous Cell Carcinoma

doi: 10.34133/cancomm.0014

Figure Lengend Snippet: CD11c mediates phosphorylated SMAD3 nuclear translocation to perform its functions. (A) Left panel, representative confocal images showing p-SMAD3 levels in ITGAX- OE KYSE30 cells treated without or with TGFBR1 inhibitor SB505124. Antibody for each channel is labeled in the panels: p-SMAD3 (green), DiI (red, for cell membrane), and DAPI (blue, for cell nuclei). Right panel, quantitative results of mean fluorescence intensity of p-SMAD3. Data represent mean ± SEM from 3 independent experiments. (B) Left panel, representative confocal images showing ITGAX- KO (sg ITGAX ) KYSE30 cells treated without or with TGFβ1. Antibody for each channel is labeled in the panels: p-SMAD3 (green), DiI (red, for cell membrane), and DAPI (blue, for cell nuclei). Right panel, quantitative results of mean fluorescence intensity of p-SMAD3. Data represent mean ± SEM from 3 independent experiments. (C) Immunoblot of the coimmunoprecipitation products collected from whole cell lysates with SMAD3 (upper panel) or CD11c antibody (lower panel) in KYSE30 cells. (D) Western blot analysis of immunoprecipitation products collected using SMAD3 antibody shows the interaction of TGFBR1-SMAD3 in KYSE30 cells with ITGAX KO or OE. (E) Coimmunoprecipitation assay of the binding of CD11c truncated mutants and SMAD3 (Flag). (F) Cell membrane (marked by Na, K-ATPase), cytoplasm (marked by β-actin), and nucleus (marked by Lamin B1) fractions of ITGAX -OE KYSE30 cells were collected and subjected to IB analysis of p-SMAD3 and SMAD3. (G) Chromatin immunoprecipitation-coupled qPCR assays show enrichment of CD80 or CD86 promoter in cell lysates obtained with anti-p-SMAD3 antibody in KYSE30 cells stimulated with vehicle (Ctrl) or TGFβ1. (H) Luciferase reporter assays in KYSE30 cells using the indicated reporter plasmids of CD80 (left) or CD86 (right) promoters or siRNA targeting SMAD3 . Data are mean ± SEM from 3 experiments, and each had 3 replicates. P values from Student t test. Abbreviations: ITGAX , integrin alpha X; sgCtrl, single guide RNA control; sg ITGAX , single guide RNA targeting ITGAX gene; Vector, control for OE group; OE, overexpression; Δ, deletion; FG-GAP (1 and 2), Phenylalanine-Glycine-GAP repeat fragment 1 to 2; FG-GAP (3 to 7), Phenylalanine-Glycine-GAP repeat fragment 3 to 7; ICD, intracellular domain; Wt- CD80 -P, wild-type CD80 promoter; Mut -CD80 -P, mutant-type CD80 promoter (without p-SMAD3 binding motif); Wt- CD86 -P, wild-type CD86 promoter; Mut- CD86 -P, mutant-type CD86 promoter (without p-SMAD3 binding motif); SMAD3, mothers against decapentaplegic homolog 3; p-SMAD3, phosphorylated SMAD3;. HA, HA tag, CD11c-Δ; Flag, Flag-SMAD3; TGFBR1, transforming growth factor beta receptor I; IB, immunoblot; IP, immunoprecipitation; TGFβ1, transforming growth factor beta 1; Ctrl, control.

Article Snippet: The protein level of TGFβ1 was analyzed using a human TGFβ1 enzyme-linked immunosorbent assay (ELISA) kit (Proteintech, Cat. #KE00002) according to the manufacturer’s instructions.

Techniques: Translocation Assay, Labeling, Membrane, Fluorescence, Western Blot, Immunoprecipitation, Co-Immunoprecipitation Assay, Binding Assay, Chromatin Immunoprecipitation, Luciferase, Control, Plasmid Preparation, Over Expression, Mutagenesis

Journal: Cancer Communications

Article Title: Ectopic CD11c Drives SMAD3-Mediated Aberrant Antigen Presentation and Epithelial–Mesenchymal Transition in Esophageal Squamous Cell Carcinoma

doi: 10.34133/cancomm.0014

Figure Lengend Snippet: Ectopic CD11c expression in epithelial cells is associated with TP53 mutations. (A) Copy number variations of the ITGAX gene in multistage human ESCC development . Each point indicates a micro-biopsy sample. Sample size at each stage: NOR, 603; LGIN, 245; HGIN, 247; and ESCC, 180. (B) Levels of ITGAX copy number (GISTIC score) among human esophageal epithelial clones with different TP53 states . Each point represents one epithelial clone. Sample size for each group: WT, 35; 1 mut, 24; >1 mut, 8; and Loss, 11. (C) Effect of TP53- knockout on ITGAX copy number in human esophageal epithelial cells. Top: Representative images of ITGAX copy number in TP53 -unknockout or TP53 -knockout HET-1A, KYSE150, and KYSE30 cells with ITGAX -specific probe (red, arrow pointed). Bottom: Quantitative statistics of the percentage of cells with ITGAX copy number in TP53 -unknockout versus TP53 -knockout cells. The number of cells ( n ) obtained from 3 independent experiments is indicated on each bar. (D) Immunoblot of CD11c in HET-1A, KYSE150, and KYSE30 cells with or without TP53 KO. Each experiment had 3 biological repeats. (E) Proposed role of ectopically expressed CD11c for cancer cells to escape immune killing and acquire malignant phenotypes. TP53 loss-associated ITGAX amplification results in CD11c ectopic overexpression in epithelial cells. CD11c interacts with SMAD3 and enhances its binding to TGFβ/TGFBR1, promoting SMAD3 phosphorylation. Hyperactivated SMAD3 subsequently translocates to the nucleus, where it suppresses CD80/CD86 transcription while up-regulating the levels of MHC class II molecules. Together, these changes impair tumor cell-mediated antigen presentation and induce EMT, thereby promoting cancer development. In (A) to (C), P values were from the Wilcoxon rank-sum test. ns, not significant. Abbreviations: ITGAX , integrin alpha X; NOR, normal epithelial tissue; INF, inflammatory tissue; LGIN, low-grade intraepithelial neoplasia tissue; HGIN, high-grade intraepithelial neoplasia tissue; ESCC, esophageal squamous cell carcinoma; GISTIC, Genomic Identification of Significant Targets in Cancer; WT, wild type; 1 mut, one mutation; >1 mut, multiple mutations; Loss, TP53 biallelic loss; Ctrl, control; EMT, epithelial–mesenchymal transition; SMAD3, mothers against decapentaplegic homolog 3; P, phosphorylation; TGFβ, transforming growth factor beta; TGFBR1, transforming growth factor beta receptor I; Treg, regulatory T cell; MHC II, major histocompatibility complex class II.

Article Snippet: The protein level of TGFβ1 was analyzed using a human TGFβ1 enzyme-linked immunosorbent assay (ELISA) kit (Proteintech, Cat. #KE00002) according to the manufacturer’s instructions.

Techniques: Expressing, Clone Assay, Knock-Out, Western Blot, Amplification, Over Expression, Binding Assay, Phospho-proteomics, Immunopeptidomics, Mutagenesis, Control

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: ADAMTS1 levels were increased in the serum of patients with myocardial fibrosis, in the hearts of mice after CFPMI and in cardiac fibroblasts stimulated with fibrotic factors. A . ADAMTS1 levels in clinical serum samples were evaluated via ELISA. N = 30. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. B . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. C . Survival rate of mice in each group. D . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. E and F . ADAMTS1 expression in mouse heart tissue was determined through Western blot and IHC. Human cardiac fibroblasts were treated with 10 ng/mL TGF-β1 or 0.1 μM Ang II for 12, 24, or 48 h to construct in vitro models. The cells were divided into four groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups. G and H . Western blot detection of the expression of ADAMTS1, Collagen I and FN. I. The expression levels of ADAMTS1, Collagen I and FN were measured via qRT‒PCR. After the optimal time point (48 h) was determined, the mice were further divided into the control, TGF-β1 and Ang II groups. J . ADAMTS1 expression in cells was assessed by IF. Human cardiac fibroblasts were treated with 10 ng/mL TGF-β1 or 0.1 μM Ang II for 15, 30, 60, 90, and 120 min. K. qRT-PCR detection of the mRNA expression of ADAMTS1, Collagen I and FN. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiac fibroblasts were incubated with 10 ng/mL TGF-β1 (HY- P78168 , MCE) or 0.1 μM Ang II (HY-13948, MCE) for 12, 24, and 48 h to establish in vitro models, and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 (50698-M08H-B, Sino Biological) for 12, 24, and 48 h to establish in vitro models (Weng et al. ), which was divided into the following groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups.

Techniques: Enzyme-linked Immunosorbent Assay, Construct, Staining, Expressing, Western Blot, In Vitro, Control, Quantitative RT-PCR

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: SMAD2 regulated ADAMTS1 expression in human and mouse cardiac fibroblasts induced by TGF-β1. We used 10 ng/mL TGF-β1 to induce human and mouse cardiac fibroblasts for 15, 30, 60, 90, and 120 min. A and B . Western blot analysis of the expression of SMAD2, p-SMAD2 and ADAMTS1. Human and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 for 48 h, and the cells were divided into Control and TGF-β1 groups. C . Human and mouse cardiac fibroblasts were induced with 10 ng/mL TGF-β1 for 12 h, 24 h, 48 h, and 72 h. qRT‒PCR was performed to detect the mRNA levels of ADAMTS1, Collagen I, and FN. D . IF staining was conducted to determine the expression of p-SMAD2 and ADAMTS1 at 48 h. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. E . p-SMAD2 expression in mouse heart tissue was measured by IHC. F . ChIP verification of the interaction between SMAD2 and ADAMTS1. Then, we interfered with SMAD2 expression. The groups were the si-NC, si-SMAD2-1, and si-SMAD2-2 groups. G . SMAD2 expression was assessed via qRT‒PCR. After the best si-SMAD2 was selected, the cells were further divided into the si-NC, TGF-β1 (48 h), si-SMAD2, and TGF-β1 + si-SMAD2 groups. H . Western blot detection of ADAMTS1, Collagen I, and FN expression in cells. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiac fibroblasts were incubated with 10 ng/mL TGF-β1 (HY- P78168 , MCE) or 0.1 μM Ang II (HY-13948, MCE) for 12, 24, and 48 h to establish in vitro models, and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 (50698-M08H-B, Sino Biological) for 12, 24, and 48 h to establish in vitro models (Weng et al. ), which was divided into the following groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups.

Techniques: Expressing, Western Blot, Control, Staining, Construct

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: Overexpression of ADAMTS1 enhanced the production of collagen fiber proteins in human and mouse cardiac fibroblasts induced by TGF-β1. We overexpressed ADAMTS1 and divided the cells into the oe-NC, TGF-β1 (48 h), oe-ADAMTS1, and TGF-β1 + oe-ADAMTS1 groups. A . Western blot detection of ADAMTS1, Collagen I, FN, and α-SMA expression. B-D . IF staining of Collagen I and FN expression. Additionally, we interfered with ADAMTS1; the cells were grouped as follows: si-NC, si-ADAMTS1-1, and si-ADAMTS1-2. E . ADAMTS1 expression was determined through qRT‒PCR. After the best si-ADAMTS1 was selected, the cells were further divided into the si-NC, TGF-β1 (48 h), si-ADAMTS1, and TGF-β1 + si-ADAMTS1 groups. F and G . Western blot detection of ADAMTS1, Collagen I, FN, and α-SMA expression. H and I . IF staining of Collagen I and FN expression. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiac fibroblasts were incubated with 10 ng/mL TGF-β1 (HY- P78168 , MCE) or 0.1 μM Ang II (HY-13948, MCE) for 12, 24, and 48 h to establish in vitro models, and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 (50698-M08H-B, Sino Biological) for 12, 24, and 48 h to establish in vitro models (Weng et al. ), which was divided into the following groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups.

Techniques: Over Expression, Western Blot, Expressing, Staining

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: Knockdown of ADAMTS1 alleviated TGF-β1-induced fibrosis by downregulating HDAC6 protein expression. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. A . HDAC6 expression in mouse heart tissue was determined through IHC. After the best si-ADAMTS1 was selected, the cells were further divided into the si-NC and si-ADAMTS1 groups. B . ADAMTS1 and HDAC6 expression was assessed via qRT‒PCR. C . Western blot analysis of ADAMTS1 and HDAC6 expression. Furthermore, we overexpressed ADAMTS1 and divided the cells into the oe-NC, TGF-β1, oe-ADAMTS1, and TGF-β1 + oe-ADAMTS1 groups. D . Western blot detection of HDAC6 expression. After the si-ADAMTS1 group was selected, the cells were further divided into the si-NC, TGF-β1, si-ADAMTS1, and TGF-β1 + si-ADAMTS1 groups. E. Western blot detection of HDAC6 expression. Moreover, cells were treated with the HDAC6 inhibitor ACY1215 (5 μM) for 48 h and then divided into the si-NC, si-ADAMTS1, si-NC + ACY1215, and si-ADAMTS1 + ACY1215 groups. F. Western blot detection of TGF-β1, Collagen I, FN, and ADAMTS1 expression. Subsequently, experiments were performed in human and mouse cardiac fibroblasts with ADAMTS1 overexpression followed by treatment with the HDAC6 inhibitor ACY1215. G. Western blot analysis was conducted to detect the expression levels of ADAMTS1 and HDAC6. H. Western blot analysis was performed to determine the expression levels of TGF-β1, Collagen I, and FN. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiac fibroblasts were incubated with 10 ng/mL TGF-β1 (HY- P78168 , MCE) or 0.1 μM Ang II (HY-13948, MCE) for 12, 24, and 48 h to establish in vitro models, and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 (50698-M08H-B, Sino Biological) for 12, 24, and 48 h to establish in vitro models (Weng et al. ), which was divided into the following groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups.

Techniques: Knockdown, Expressing, Construct, Western Blot, Over Expression

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: ADAMTS1 interacted with HDAC6 during fibrosis. A . Co-IP verification of the interaction of ADAMTS1 with HDAC6, with ADAMTS1 as the bait protein. In the experiment, we first captured the ADAMTS1 protein using a specific antibody and then used Co-IP technology to detect its interaction with HDAC6. B . Co-IP verification of the interaction of HDAC6 with ADAMTS1, with HDAC6 as the bait protein. In the experiment, we captured the HDAC6 protein using a specific antibody and then used Co-IP technology to detect its interaction with ADAMTS1. We used 10 ng/mL TGF-β1 to induce human and mouse cardiac fibroblasts for 48 h, and the cells were further divided into control and TGF-β1 groups. C . IF staining was performed to evaluate the colocalization of ADAMTS1 and HDAC6 in TGF-β1-treated human and mouse cardiac fibroblasts. D . Changes in the level of ubiquitinated HDAC6 protein in human and mouse cardiac fibroblasts transfected with oe-ADAMTS1/si-ADAMTS1 or oe-NC/si-NC in the presence of 10 μM MG132. E. In si-ADAMTS1-treated human and mouse cardiac fibroblasts, MG132 (10 μM) was added to detect the ubiquitination levels of ADAMTS1 protein. F. Human and mouse cardiac fibroblasts were treated with TGF-β1 and subjected to SMAD2 knockdown to determine the ubiquitination levels of ADAMTS1 protein. N = 3. *** P < 0.001

Article Snippet: Human cardiac fibroblasts were incubated with 10 ng/mL TGF-β1 (HY- P78168 , MCE) or 0.1 μM Ang II (HY-13948, MCE) for 12, 24, and 48 h to establish in vitro models, and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 (50698-M08H-B, Sino Biological) for 12, 24, and 48 h to establish in vitro models (Weng et al. ), which was divided into the following groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups.

Techniques: Co-Immunoprecipitation Assay, Control, Staining, Transfection, Ubiquitin Proteomics, Knockdown

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: AAV-shRNA-ADAMTS1 treatment alleviated myocardial fibrosis and improved cardiac function after CFPMI. The mice were randomly divided into the Sham, CFPMI, and CFPMI + sh-ADAMTS1 groups. A . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. B . Survival rate of mice in each group. C . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. D . HDAC6 expression was determined through qRT‒PCR. E. Western blot analysis of the expression of TGF-β1, SMAD2, and p-SMAD2 in mouse heart tissue. F. Western blot detection of Collagen I, and FN expression. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiac fibroblasts were incubated with 10 ng/mL TGF-β1 (HY- P78168 , MCE) or 0.1 μM Ang II (HY-13948, MCE) for 12, 24, and 48 h to establish in vitro models, and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 (50698-M08H-B, Sino Biological) for 12, 24, and 48 h to establish in vitro models (Weng et al. ), which was divided into the following groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups.

Techniques: shRNA, Staining, Expressing, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: AAV-shRNA-HDAC6 transfection combined with ADAMTS1 inhibitor treatment alleviated myocardial fibrosis and improved cardiac function after CFPMI. The mice were randomly divided into the Sham, CFPMI, CFPMI + anti-ADAMTS1, CFPMI + sh-HDAC6, and CFPMI + anti-ADAMTS1 + sh-HDAC6 groups. A . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. B . Survival rate of mice in each group. C and D . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. E. HDAC6 expression was determined through qRT‒PCR. F. Western blot analysis of the expression of TGF-β1, SMAD2, and p-SMAD2 in mouse heart tissue. G. Western blot detection of Collagen I and FN expression. N = 3. ** P < 0.01, *** P < 0.001

Article Snippet: Human cardiac fibroblasts were incubated with 10 ng/mL TGF-β1 (HY- P78168 , MCE) or 0.1 μM Ang II (HY-13948, MCE) for 12, 24, and 48 h to establish in vitro models, and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 (50698-M08H-B, Sino Biological) for 12, 24, and 48 h to establish in vitro models (Weng et al. ), which was divided into the following groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups.

Techniques: shRNA, Transfection, Staining, Expressing, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: TGF-β1/SMAD2 regulated ADAMTS1 by mediating CFPMI through HDAC6 ubiquitination

Article Snippet: Human cardiac fibroblasts were incubated with 10 ng/mL TGF-β1 (HY- P78168 , MCE) or 0.1 μM Ang II (HY-13948, MCE) for 12, 24, and 48 h to establish in vitro models, and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 (50698-M08H-B, Sino Biological) for 12, 24, and 48 h to establish in vitro models (Weng et al. ), which was divided into the following groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups.

Techniques: Ubiquitin Proteomics

Journal: Reviews in Cardiovascular Medicine

Article Title: Targeting Tumor Necrosis Factor-α Mitigates Glucose Fluctuation-Induced Aortic Valve Fibrosis: Insights From Diabetic Rat Models

doi: 10.31083/RCM42804

Figure Lengend Snippet: TNF- α -mediated inflammation could exacerbate fibrosis in vitro in porcine aortic valve interstitial cells across different glucose levels . (A,B,E,F) In the primary porcine aortic valve interstitial cells (pAVICs) groups, both HG and GF conditions were found to upregulate collagen 1 protein expression. Moreover, the proinflammatory cytokine TNF- α aggravated this upregulation under HG conditions, with an even more pronounced effect observed under GF conditions. Conversely, the inhibition of TNF- α could reverse these upregulations in both the HG and GF groups. (C,D,G,H) TGF- β 1 protein expression increased under both HG and GF conditions, with TNF- α further enhancing this increase, especially under GF conditions. The inhibition of TNF- α reversed these effects in both groups (n = 4 per group). The data are presented as the mean ± SEM. Statistical analyses were conducted utilizing one-way ANOVA, followed by post hoc corrections to account for multiple comparisons (Fig. E–H). * p < 0.05.

Article Snippet: In addition, the concentrated SABC-POD (Mouse/Rabbit IgG) kit (SA2010, BOSTER Biological Technology Co., Ltd., Wuhan, China), DyLight 488 conjugated AffiniPure goat anti-mouse IgG (H + L) (BA1126, BOSTER Biological Technology Co., Ltd., Wuhan, China), DyLight 594 conjugated AffiniPure goat anti-rabbit IgG (H + L) (BA1142, BOSTER Biological Technology Co., Ltd., Wuhan, China), ethylenediaminetetraacetic acid (EDTA) antigen retrieval solution (AR0023, BOSTER Biological Technology Co., Ltd., Wuhan, China), 4 ′ ,6-diamidino-2-phenylindole (DAPI) staining solution (AR1176, BOSTER Biological Technology Co., Ltd., Wuhan, China), human TNF- α enzyme-linked immunosorbent assay (ELISA) kit (EK0525, BOSTER Biological Technology Co., Ltd., Wuhan, China), and human TGF- β 1 ELISA kit (EK0513, BOSTER Biological Technology Co., Ltd., Wuhan, China) were obtained from Boster, China.

Techniques: In Vitro, Expressing, Inhibition