Journal: Molecular Medicine

Article Title: Inhibition of tartrate-resistant acid phosphatase 5 can prevent cardiac fibrosis after myocardial infarction

doi: 10.1186/s10020-024-00856-1

Figure Lengend Snippet: Inhibition of ACP5 suppresses CF proliferation, migration, and transition into myofibroblasts. ( A ) Relative mRNA expression levels after ACP5 silencing ( n = 3/group). ( B - C ) Western blot analysis of the silencing efficiency of ACP5 ( n = 3/group). ( D - E ) EdU assay was used to detect the cell proliferation rate of CFs ( n = 4/group); scale bar = 100 μm. ( F ) OD values of CFs in different groups ( n = 4/group). ( G - H ) Cell migration was assessed by a wound-healing assay ( n = 4/group); scale bar = 100 μm. ( I - J ) Cell migration was assessed by Transwell assays ( n = 4/group); scale bar = 100 μm. ( K - N ) Relative mRNA expression levels of ACP5, α-SMA, COL1, and COL3 in CFs in different groups ( n ≥ 3/group). ( O - Q ) Western blot analysis of α-SMA and COL1 ( n = 3/group). * P < 0.05, ** P < 0.01, *** P < 0.001, and ns: P > 0.05

Article Snippet: The Mouse ACP5 ELISA Kit was purchased from Signalway Antibody (USA, #EK12408), and the Human ACP5 ELISA Kit was purchased from BOSTER (China, #EK2138).

Techniques: Inhibition, Migration, Expressing, Western Blot, EdU Assay, Wound Healing Assay

Journal: Molecular Medicine

Article Title: Inhibition of tartrate-resistant acid phosphatase 5 can prevent cardiac fibrosis after myocardial infarction

doi: 10.1186/s10020-024-00856-1

Figure Lengend Snippet: Overexpression of ACP5 promotes CF proliferation, migration, and transition into myofibroblasts. ( A ) Relative mRNA expression levels of ACP5 and the overexpression efficiency ( n = 4/group). ( B - C ) Western blot analysis of the overexpression efficiency of ACP5 ( n = 3/group). ( D - E ) The EdU assay was used to detect the cell proliferation rate of CFs ( n = 4/group); scale bar = 100 μm. ( F ) OD values of CFs in different groups ( n = 3/group). ( G - H ) Cell migration was assessed by a wound-healing assay ( n = 4/group); scale bar = 100 μm. ( I - J ) Cell migration was assessed by Transwell assays ( n = 4/group); scale bar = 100 μm. ( K - N ) Relative mRNA expression levels of ACP5, α-SMA, COL1, and COL3 in CFs in different groups ( n = 4/group). ( O - Q ) Western blot analysis of α-SMA and COL1 ( n = 3/group). * P < 0.05, ** P < 0.01, *** P < 0.001, and ns: P > 0.05

Article Snippet: The Mouse ACP5 ELISA Kit was purchased from Signalway Antibody (USA, #EK12408), and the Human ACP5 ELISA Kit was purchased from BOSTER (China, #EK2138).

Techniques: Over Expression, Migration, Expressing, Western Blot, EdU Assay, Wound Healing Assay

Journal: Molecular Medicine

Article Title: Inhibition of tartrate-resistant acid phosphatase 5 can prevent cardiac fibrosis after myocardial infarction

doi: 10.1186/s10020-024-00856-1

Figure Lengend Snippet: ACP5 inhibition suppresses fibrosis in MI mice and enhances cardiac function. ( A ) Masson staining and Sirius red staining in each group ( n = 6/group; upper layer, scale bar = 1000 μm; middle layer, scale bar = 100 μm; lower layer, scale bar = 100 μm). ( B ) Masson staining of fibrosis. ( C ) Sirius red staining of the collagen area. ( D ) Representative images of echocardiography. ( E - F ) Echocardiographic measurements of LVEF (E) and LVFS (F) ( n = 6/group). ( G - I ) Western blot analysis of α-SMA and COL1 in the hearts of mice in different groups ( n = 3/group). ( J - K ) Immunofluorescence staining of α-SMA in the hearts of mice in different groups ( n = 4/group); scale bar = 50 μm. (The bottom-layer image in Fig. 4J is the locally enlarged image circled in the Merge graph, bar = 20 μm). * P < 0.05, ** P < 0.01, *** P < 0.001, and ns: P > 0.05

Article Snippet: The Mouse ACP5 ELISA Kit was purchased from Signalway Antibody (USA, #EK12408), and the Human ACP5 ELISA Kit was purchased from BOSTER (China, #EK2138).

Techniques: Inhibition, Staining, Western Blot, Immunofluorescence

Journal: Molecular Medicine

Article Title: Inhibition of tartrate-resistant acid phosphatase 5 can prevent cardiac fibrosis after myocardial infarction

doi: 10.1186/s10020-024-00856-1

Figure Lengend Snippet: ACP5 affects the GSK3β/β-catenin signal transduction pathway. ( A - D ) Western blot analysis of the expression levels of ACP5, p-GSK3β, GSK3β, and β-catenin in ACP5-deficient CFs ( n = 3/group). ( E - H ) Western blot analysis of the expression levels of ACP5, p-GSK3β, GSK3β, and β-catenin in CFs overexpressing ACP5 ( n = 3/group). ( I - L ) Western blot analysis of the expression levels of ACP5, p-GSK3β, GSK3β, and β-catenin in the hearts of mice in different groups ( n ≥ 3/group). * P < 0.05, ** P < 0.01, *** P < 0.001, and ns: P > 0.05

Article Snippet: The Mouse ACP5 ELISA Kit was purchased from Signalway Antibody (USA, #EK12408), and the Human ACP5 ELISA Kit was purchased from BOSTER (China, #EK2138).

Techniques: Transduction, Western Blot, Expressing

Journal: Molecular Medicine

Article Title: Inhibition of tartrate-resistant acid phosphatase 5 can prevent cardiac fibrosis after myocardial infarction

doi: 10.1186/s10020-024-00856-1

Figure Lengend Snippet: ACP5 affects CF activation by regulating ERK. ( A - B ) Western blot analysis of the expression levels of p-ERK and ERK in ACP5-deficient CFs ( n = 3/group). ( C - D ) Western blot analysis of the expression levels of p-ERK and ERK in CFs overexpressing ACP5 ( n = 3/group). ( E - F ) Western blot analysis of the expression levels of p-ERK and ERK in the hearts of mice in different groups ( n = 3/group). ( G - K ) Western blot analysis of the expression levels of ACP5, p-ERK, ERK, p-GSK3β, GSK3β, and β-catenin in CFs pretreated with Ro 67-7476 (an ERK agonist) ( n = 3/group). ( L - M ) The EdU assay was used to detect the proliferation rate of CFs pretreated with Ro 67-7476 (an ERK agonist) ( n = 4/group); scale bar = 100 μm. ( N - O ) Cell migration was assessed by Transwell assays ( n = 4/group); scale bar = 100 μm. ( P - R ) Western blot analysis of α-SMA and COL1 in the hearts of mice in different groups ( n = 3/group).* P < 0.05, ** P < 0.01, *** P < 0.001, and ns: P > 0.05

Article Snippet: The Mouse ACP5 ELISA Kit was purchased from Signalway Antibody (USA, #EK12408), and the Human ACP5 ELISA Kit was purchased from BOSTER (China, #EK2138).

Techniques: Activation Assay, Western Blot, Expressing, EdU Assay, Migration

Journal: Molecular Medicine

Article Title: Inhibition of tartrate-resistant acid phosphatase 5 can prevent cardiac fibrosis after myocardial infarction

doi: 10.1186/s10020-024-00856-1

Figure Lengend Snippet: Schematic diagram of the mechanism of ACP5 in myocardial fibrosis after MI. Under the stimulation of MI or Ang II, the increased expression of ACP5 activates the ERK/GSK3β/β-catenin signaling pathway, which promotes the transformation of CFs into myofibroblasts with more active proliferation, migration and fibrosis, leading to the onset of myocardial fibrosis. (Generated by Figdraw)

Article Snippet: The Mouse ACP5 ELISA Kit was purchased from Signalway Antibody (USA, #EK12408), and the Human ACP5 ELISA Kit was purchased from BOSTER (China, #EK2138).

Techniques: Expressing, Transformation Assay, Migration, Generated

Journal: Mediators of Inflammation

Article Title: Monocytes Undergo Functional Reprogramming to Generate Immunosuppression through HIF-1 α Signaling Pathway in the Late Phase of Sepsis

doi: 10.1155/2020/4235909

Figure Lengend Snippet: Its cytokine production in the blood serum. (a) In the clinical study, the higher levels of TNF- α , IL-1 β , IL-6, IL-18, CCL3, and CCL5 except for IL-10 in the early phase of sepsis compared with the late phase of sepsis and the healthy participants, n = 30. ∗∗ P < 0.01 represented the significant difference; ## P < 0.01: compared with the healthy participants, the difference was of significance. (b) Ade-HIF-1 α injection increased the production of TNF- α , IL-1 β , IL-6, IL-18, CCL3, and CCL5 except for IL-10 in the late stage of the sepsis rat model. ∗∗ P < 0.01: compared with the normal control group, the difference was of significance; # P < 0.05: compared with the sepsis rat model group and the adenovirus control group; & P < 0.05: compared with the sepsis rat model group. Besides, data were presented as mean ± standard deviation, and data was repeated in triplicate.

Article Snippet: Then, the serum of the blood samples was dividedly analyzed with ELISA assay kits as per the manufacturer's protocol to quantify the concentration of IL-1 β (catalog:70-EK101B-96, homo; catalog:70-EK201B/3-96, mus; MultiSciences, China), IL-18 (catalog: 70-EK118-48, homo; catalog: 70-EK218-96, mus; MultiSciences, China), IL-6 (catalog: 70-EK106/2-96, homo; catalog: 70-EK206/3-96, mus; MultiSciences, China), IL-10 (catalog: 70-EK110/2-96, homo; catalog: 70-EK210/3-96, mus; MultiSciences, China), TNF- α (catalog: 70-EK182-96, homo; catalog: 70-EK282/3-96, mus; MultiSciences, China), CCL3 (catalog: 70-EK161-96, homo; catalog: 70-EK261/2-96, mus; MultiSciences, China), and CCL5 (catalog: 70-EK1129-96, homo; catalog: 70-EK2129/2-96, mus; MultiSciences, China) in triplicates.

Techniques: Injection, Standard Deviation

Journal: Theranostics

Article Title: Fibrinogen-like protein 2 aggravates nonalcoholic steatohepatitis via interaction with TLR4, eliciting inflammation in macrophages and inducing hepatic lipid metabolism disorder

doi: 10.7150/thno.44297

Figure Lengend Snippet: Fgl2 deficiency attenuated liver inflammatory injury in NASH mice. In MCD-fed or HFD-fed WT and fgl2-/- mice, HE staining was performed to detect histological changes in the liver (A). The NAFLD activity score was evaluated (B). BMDMs were isolated from WT or fgl2-/- mice and injected into macrophage-depleted WT NASH mice (C). Histological changes were detected by HE staining (D, arrows indicate inflammatory infiltration). Serum ALT, AST, LDH (E) and fasting glucose (F) were tested by an automatic biochemical analyzer (n=10 in each group). The levels of serum insulin were tested by an ELISA kit (F). The levels of the proinflammatory cytokines TNF-α, MCP-1, IL-6, IL-1β and IL-18 in the liver were tested by ELISAs (G). For bar graphs, n=6-10 in each group. The data represent the mean ± SD from at least three independent experiments. Statistical differences were determined by two-way ANOVA. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; ns, not significant.

Article Snippet: Cytokines in liver homogenates from the mice and the culture supernatants of BMDMs were determined by ELISA kits (TNF-α 1217202, IL-1β 1210122, MCP-11217392, IL-6 1210602, Dakewe Biotech, China; IL-18 70-EK218-96, MutiSciences, China) according to the manufacturers' instructions.

Techniques: Staining, Activity Assay, Isolation, Injection, Enzyme-linked Immunosorbent Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Salidroside Attenuates High-Fat Diet-Induced Nonalcoholic Fatty Liver Disease via AMPK-Dependent TXNIP/NLRP3 Pathway

doi: 10.1155/2018/8597897

Figure Lengend Snippet: Salidroside (SAL) improves insulin sensitivity and suppresses NLRP3 inflammasome activation in hepatocytes. After cultured overnight in the serum-free medium which contains normal glucose (NG, 5.5 mM), primary mouse hepatocytes were incubated in the serum-free medium which contains 30 mM glucose and 100 nM insulin (HG) and treated with 10 μ M SAL for the indicated periods of time (0–72 h), or treated with SAL (0.1, 1, and 10 μ M) for 72 h. For assessment of insulin sensitivity in vitro , hepatocytes were treated as indicated, the medium was removed, and cells were incubated in fresh serum-free DMEM containing insulin (10 nM) for 20 min before protein samples were extracted. Protein sample were extracted from hepatocytes or supernatant (SN). The phosphorylation of Akt and GSK3 β (a, d) and the activation of NLRP3 inflammasome (c, f) were analyzed by immunoblot. The supernatant IL-1 β concentration (b, e) was measured by ELISA method. † P < 0.05, †† P < 0.01 versus NG; ∗ P < 0.05, ∗∗ P < 0.01 versus HG. Values are means ± s.e.m. ((a), (c), (d), and (f): n = 4; (b) and (e): n = 3).

Article Snippet: The serum IL-1 β level was measured using an ELISA kit (number 70-EK2195) purchased from MultiSciences Biotech (Hangzhou, China) according to the manufacturer's instructions.

Techniques: Activation Assay, Cell Culture, Incubation, In Vitro, Phospho-proteomics, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Salidroside Attenuates High-Fat Diet-Induced Nonalcoholic Fatty Liver Disease via AMPK-Dependent TXNIP/NLRP3 Pathway

doi: 10.1155/2018/8597897

Figure Lengend Snippet: Salidroside (SAL) protects hepatocytes against ROS overproduction. After cultured overnight in the serum-free medium which contains normal glucose (NG, 5.5 mM), hepatocytes were incubated in the serum-free medium which contains 30 mM glucose and 100 nM insulin (HG) and treated with 10 μ M SAL for the indicated periods of time (0–72 h), or treated with SAL (0.1, 1, 10 μ M) and N-acetylcysteine (NAC, 5 mM) for 72 h. The ROS levels were detected using DCF-DA and MitoSOX as indicated (a–d). The Oil Red O staining (e) and lipid content analysis (f) were performed. Scale bar = 200 μ m. Protein sample was extracted from hepatocytes or supernatant (SN). For assessment of insulin sensitivity in vitro , hepatocytes were treated as indicated, the medium was removed, and cells were incubated in fresh serum-free DMEM containing insulin (10 nM) for 20 min before protein samples were extracted. The phosphorylation of AMPK, ACC (g), Akt, and GSK3 β (h) and the activation of NLRP3 inflammasome (j) were analyzed by immunoblot. The supernatant IL-1 β concentration was measured by ELISA method (i). † P < 0.05, †† P < 0.01 versus NG; ∗ P < 0.05, ∗∗ P < 0.01 versus HG. Values are means ± s.e.m. ((a)–(d), (f)–(h), and (j): n = 4; (e) and (i): n = 3).

Article Snippet: The serum IL-1 β level was measured using an ELISA kit (number 70-EK2195) purchased from MultiSciences Biotech (Hangzhou, China) according to the manufacturer's instructions.

Techniques: Cell Culture, Incubation, Staining, In Vitro, Phospho-proteomics, Activation Assay, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay