cd36 overexpression  (Addgene inc)

Journal: Frontiers in Pharmacology

Article Title: Astragaloside IV attenuates renal tubule injury in DKD rats via suppression of CD36-mediated NLRP3 inflammasome activation

doi: 10.3389/fphar.2024.1285797

Figure Lengend Snippet: Primer sequences.

Article Snippet: The CD36 overexpression adenovirus was acquired from Genechem Co., Ltd. (Shanghai, China).

Techniques: Sequencing

Journal: Frontiers in Pharmacology

Article Title: Astragaloside IV attenuates renal tubule injury in DKD rats via suppression of CD36-mediated NLRP3 inflammasome activation

doi: 10.3389/fphar.2024.1285797

Figure Lengend Snippet: AS-IV inhibits CD36 production and NLRP3 inflammasome activation in DKD rats. (A) Western blot analysis of ASC, Cleaved-Caspase-1, NLRP3, Cleaved-IL-1β, CD36, GSDMD-D, and Cleaved-GSDMD-D. (B–H) Protein levels of ASC, Cleaved-Caspase-1, NLRP3, Cleaved-IL-1β, and CD36 after normalization to α-tubulin, protein levels of GSDMD-D, and Cleaved-GSDMD-D after normalization to GAPDH. All data are presented as the mean ± SD ( n = 6). ## p < 0.01 vs. Control; * p < 0.05 and ** p < 0.01 vs. Model.

Article Snippet: The CD36 overexpression adenovirus was acquired from Genechem Co., Ltd. (Shanghai, China).

Techniques: Activation Assay, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Astragaloside IV attenuates renal tubule injury in DKD rats via suppression of CD36-mediated NLRP3 inflammasome activation

doi: 10.3389/fphar.2024.1285797

Figure Lengend Snippet: AS-IV inhibits CD36 expression, lipid accumulation, and lipid ROS production in PA-induced HK-2 cells. (B–D) The mRNA expression levels of CD36, CPT1, and PPAP-α were determined by RT-qPCR. (E,F) The protein levels of CD36 were determined by western blot and normalized to α-tubulin. (G,H) Lipid ROS production was detected by flow cytometric analysis. (I) Lipid accumulation in each group was assessed by Oil Red O (ORO) staining under light microscopy (bar = 40 μm). (J) Densitometric analysis of ORO staining. Data are expressed as the mean ± SD. ## p < 0.01 vs. Control; * p < 0.05 and ** p < 0.01 vs. PA.

Article Snippet: The CD36 overexpression adenovirus was acquired from Genechem Co., Ltd. (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Staining, Light Microscopy

Journal: Frontiers in Pharmacology

Article Title: Astragaloside IV attenuates renal tubule injury in DKD rats via suppression of CD36-mediated NLRP3 inflammasome activation

doi: 10.3389/fphar.2024.1285797

Figure Lengend Snippet: Overexpression of CD36 promotes PA-induced lipid accumulation, lipid ROS production, and activation of the NLRP3 inflammatory body and reverses the protective effect of AS-IV on the expression of lipid peroxidation-related proteins and the NLRP3 inflammatory body in PA-induced HK-2 cells. (A–G) The mRNA expression levels of PPAP-α, CPT1, CD36, ASC, Caspase-1, NLRP3, and IL-1β were determined by RT-qPCR. (H–R) Western blot analysis was conducted to determine the expression of CD36, ASC, Cleaved-Caspase-1, NLRP3, Cleaved-IL-1β, GSDMD-D, Cleaved-GSDMD-D, NGAL, KIM-1 and L-FABP. (S) Lipid ROS production was detected by flow cytometric analysis. (T) Lipid accumulation was assessed by ORO staining under light microscopy (bar = 40 μm). Data are expressed as the mean ± SD. ## p < 0.01 vs. Control; * p < 0.05 and ** p < 0.01 vs. PA; △ p < 0.05 vs. PA + AS-IV 80.

Article Snippet: The CD36 overexpression adenovirus was acquired from Genechem Co., Ltd. (Shanghai, China).

Techniques: Over Expression, Activation Assay, Expressing, Quantitative RT-PCR, Western Blot, Staining, Light Microscopy

Journal: Nature communications

Article Title: Endothelial cell CD36 regulates membrane ceramide formation, exosome fatty acid transfer and circulating fatty acid levels.

doi: 10.1038/s41467-023-39752-3

Figure Lengend Snippet: Fig. 10 | Schematic of the EC FA export pathway. A The polarized ECs control tissue FA uptake by transcytosis of circulating FAs and their export in small extracellular vesicles (sEVs) to parenchymal cells. FA binding to CD36 recruits Src to phosphorylate Cav-1 at tyrosine14, which disrupts the caveolae coat. Associated activation of nSMase2 generates the destabilizing ceramides in the caveolae. Both events initiate caveolae fission from the membrane. The caveolae internalize into small ceramide-rich intracellular vesicles (IVs) thatare then sorted into the lumen of multivesicular bodies (MVBs). The MVBs fuse with the basolateral membrane to release exosome-like small sEVs in the sub-endothelial space. The sEVs export FA and other cargo to parenchymal cells. B Possible alternate fate of endocytosed vesicles in parenchymal cells. The IVs generated in response to FA are likely to be heterogenous and a subset possibly less enriched in specific ceramides might during traffic or sorting associate with proteins such as FABPs that help target the IVs to cellular organelles instead of for secretion as sEVs. This latter fate might predominate in myocytes, cardiomyocytes, and adipocytes where most FAs are kept for local use or storage.

Article Snippet: For CD36 overexpression, mEmerald-CD36 (Addgene, #54030) and mCherry-Cav-1 (Addgene, #55008) were transfected with Lipofectamaine LTX (ThermoFisher Scientific).

Techniques: Control, Binding Assay, Activation Assay, Membrane, Generated

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ficus hirta Vahl. Ameliorates Nonalcoholic Fatty Liver Disease through Regulating Lipid Metabolism and Gut Microbiota

doi: 10.1155/2022/3474723

Figure Lengend Snippet: Amelioration of lipid accumulation in PA-induced HepG2 cells by FV. (a) Oil red O was used to measure the level of lipid accumulation (magnification 100x, scale bar = 250 μ m). (b) The oil red O-positive area was analyzed and quantified. (c–j) The relative mRNA expression levels of FABP1 , SCD1 , CD36 , HMGCR , ACACA , CCL5 , IL-1β , and TNF-α were determined by qRT-PCR. (k) The proinflammatory factor protein levels of IL-1 β , IL-6, and TNF- α . (l) The protein levels related to lipid metabolism of SREBP-1, ACOX1, CD36, CPT1 α , and HMGCR were analyzed by western blotting, and the relative ratios were calculated and expressed as the mean ± SD; n = 3. # P < 0.05 means that the difference between the NC group and the PA group is significant. ∗ P < 0.05 means that the difference between the PA group and the FV (30 mg/mL) group or the FV (15 mg/mL) group is significant.

Article Snippet: The plasmid overexpressing CD36 was designed and built by iGene Biotechnology Co., Ltd. (Guangzhou, China) to generate a CD36-overexpressed (CD36 OE) cell line, and an empty vector was used as a control. siCD36 was synthesized by RIBOBIO Co., Ltd. (Guangzhou, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ficus hirta Vahl. Ameliorates Nonalcoholic Fatty Liver Disease through Regulating Lipid Metabolism and Gut Microbiota

doi: 10.1155/2022/3474723

Figure Lengend Snippet: Effects of FV on liver lipogenesis-related markers in the mice fed with HFD. (a–h) The relative mRNA expression levels of Acaca , Cd 36, Cpt 1 α , Srebp- 1, Hmgcr , Pparα , Pparγ , and Scd 1 were determined by qRT-PCR. (i) The lipid metabolism relevant protein levels of SREBP-1, ACOX1, CD36, CPT1 α , and HMGCR were analyzed by western blotting, and the relative ratios were calculated. (j–l) Serum IL-6, IL-1 β , and TNF- α were identified by ELISA kits. (m, n) The relative mRNA expression levels of IL-1 β , TNF- α , and Ccl5 were identified by qRT-PCR. (p) The proinflammatory factor protein levels of IL-1 β , IL-6, and TNF- α were analyzed by western blotting, and their relative ratios were calculated. The densitometry was obtained by averaging repeated experiments results, normalized to GAPDH. The data are expressed as the mean ± SD, n = 3. # P < 0.05 means there is significant difference between the NFD group and the HFD group. ∗ P < 0.05 means the difference between the HFD group and the FV-L group or the FV-H group is significant.

Article Snippet: The plasmid overexpressing CD36 was designed and built by iGene Biotechnology Co., Ltd. (Guangzhou, China) to generate a CD36-overexpressed (CD36 OE) cell line, and an empty vector was used as a control. siCD36 was synthesized by RIBOBIO Co., Ltd. (Guangzhou, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ficus hirta Vahl. Ameliorates Nonalcoholic Fatty Liver Disease through Regulating Lipid Metabolism and Gut Microbiota

doi: 10.1155/2022/3474723

Figure Lengend Snippet: Alleviation of lipid accumulation and inflammation by FV through regulating CD36 in HepG2 cells with PA inducement. (a, c) CD36 protein levels after CD36 knockdown or CD36 overexpression and quantitative analysis. (b, d) TG contents of HepG2 cells in different treatment groups. (e, f) Oil red O to examine the accumulation level of lipid in HepG2 cells and their quantified scores (magnification 100x, scale bar = 250 μ m). (g) The mRNA levels of SCD1, PPAR γ , ACACA, SREBP-1, IL-1 β , and CCL5 in siCD36 HepG2 cells with or without FV administration under the condition of PA inducement. (h) The mRNA levels of PPAR γ , SREBP-1, IL-1 β , and TNF- α in CD36 OE HepG2 cells with or without FV administration under the condition of PA inducement. (i) The protein levels of CD36, HMGCR, ACOX1, and SREBP-1 in siCD36 HepG2 cells. (j) The protein levels of CD36, HMGCR, ACOX1, and SREBP-1 in CD36 OE HepG2 cells. n = 3, ∗ P < 0.05 vs. NC group for (a, c); # P < 0.05 vs. NC group, ∗ P < 0.05 vs. PA group or PA+siCD36 group/PA+CD36 OE group (b–j).

Article Snippet: The plasmid overexpressing CD36 was designed and built by iGene Biotechnology Co., Ltd. (Guangzhou, China) to generate a CD36-overexpressed (CD36 OE) cell line, and an empty vector was used as a control. siCD36 was synthesized by RIBOBIO Co., Ltd. (Guangzhou, China).

Techniques: Knockdown, Over Expression

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ficus hirta Vahl. Ameliorates Nonalcoholic Fatty Liver Disease through Regulating Lipid Metabolism and Gut Microbiota

doi: 10.1155/2022/3474723

Figure Lengend Snippet: The speculative mechanism of Ficus hirta Vahl. for treatment of NAFLD in HFD-fed mice. FV alleviates nonalcoholic fatty liver disease through adjusting lipid metabolism and inflammation via downregulation of CD36 and regulating the structure of gut microbiota.

Article Snippet: The plasmid overexpressing CD36 was designed and built by iGene Biotechnology Co., Ltd. (Guangzhou, China) to generate a CD36-overexpressed (CD36 OE) cell line, and an empty vector was used as a control. siCD36 was synthesized by RIBOBIO Co., Ltd. (Guangzhou, China).

Techniques:

Journal: Redox Biology

Article Title: Pharmaceutical targeting of succinate dehydrogenase in fibroblasts controls bleomycin-induced lung fibrosis

doi: 10.1016/j.redox.2021.102082

Figure Lengend Snippet: Perturbation of glycolysis and FAO affects fibroblast-to-myofibroblast transition. A Cellular map of metabolic enzymes and pathways targeted by drugs. B Inhibiting glycolysis downregulates profibrotic mRNA and protein levels in primary lung fibroblasts. qRT-PCR and Western blot analysis of profibrotic genes and protein (n = 3). C Representative image showing α-SMA (green) and nuclei (blue) in primary lung fibroblasts with indicated treatment for 24 h. D Inhibiting FAO upregulates profibrotic mRNA and protein levels in primary lung fibroblasts. Relative mRNA and protein levels of profibrotic markers in primary lung fibroblasts treated with etomoxir determined by qRT-PCR and Western blot (n = 3). E Relative mRNA and protein levels related to fibrosis in primary lung fibroblast treated with or without TGF-β1 and fenofibrate (n = 3). F Representative expressions of profibrotic proteins in primary lung fibroblast with indicated treatment. G Staining of lipid droplets using LipidTOX (green) in control HFL1 or cells overexpressed CD36, nuclei were counterstained with DAPI (blue). H Relative mRNA expression of transcripts related to fibrosis in control HFL1 or cells overexpressed CD36 (n = 3). Drug doses: TGF-β1 (5 ng/ml), BAY-876 (2.5 μM), 3-PO (5 mM), 2-DG (5 mM), etomoxir (10 μM), fenofibrate (1 μM). Bars represent 100 μm (C) , 10 μm (G) . The results are presented as the means ± S.D. (*p < 0.05, **P < 0.01; D and H, student's t-test; B and E, One-way ANOVA). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: CD36 overexpressed lentiviral vectors and control lentiviral vectors were obtained from Genechem and transfected into HFL1 following the manufacturer's instructions.

Techniques: Quantitative RT-PCR, Western Blot, Staining, Control, Expressing

Journal: Biochemical and biophysical research communications

Article Title: Overexpression of CD36 in mammary fibroblasts suppresses colony growth in breast cancer cell lines

doi: 10.1016/j.bbrc.2020.03.061

Figure Lengend Snippet: Activin A downregulated CD36 expression in primary fibroblasts as monitored with immunofluorescent wide-field microscopy. (a) CD36 expression in transfected primary fibroblasts was monitored with a wide-field fluorescent microscope, where nuclei were labeled with DAPI and delineated computationally in yellow. (b) CD36 expression of primary fibroblasts, cocultured with MDA-MB-231 cells, was quantitatively analyzed. Controls were primary, CD36+-, CD36−-, primary fibroblasts, and fibroblast with MDA-MB-231 coculture, supplemented with activin A neutralizing antibody. (c) CD36 expression of primary fibroblasts, in response to different concentrations of activin A at 0, 1, 2.25, and 20 ng/mL and the conditioned media from MDA-MB-231 conditioned medium were quantified. Controls were CD36+-, CD36−-, primary fibroblasts and the conditioned media supplemented with the activin A neutralizing antibody.

Article Snippet: CD36 transfection and validation Fibroblasts were transfected with CD36 overexpression and knockdown constructs using CRISPR-cas9 technology (Santa Cruz Biotechnology SC400233-ACT/KO) and validated using fluorescent microscopy per vendor’s protocol.

Techniques: Expressing, Microscopy, Transfection, Labeling

Journal: Biochemical and biophysical research communications

Article Title: Overexpression of CD36 in mammary fibroblasts suppresses colony growth in breast cancer cell lines

doi: 10.1016/j.bbrc.2020.03.061

Figure Lengend Snippet: The total production of activin A, in the co-culture assay, was reduced as a result of co-culturing cancer cell lines with the CD36+ fibroblasts. Conditioned medium was collected on (a) day 3 and (b) day 6 and quantified with the ELISA assay. MCF10A is a non-malignant cell line that was used as a reference signal. (c) Production of activin A in monocultures of CD36− fibroblast cells was increased compared to CD36+ fibroblasts.

Article Snippet: CD36 transfection and validation Fibroblasts were transfected with CD36 overexpression and knockdown constructs using CRISPR-cas9 technology (Santa Cruz Biotechnology SC400233-ACT/KO) and validated using fluorescent microscopy per vendor’s protocol.

Techniques: Co-culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Biochemical and biophysical research communications

Article Title: Overexpression of CD36 in mammary fibroblasts suppresses colony growth in breast cancer cell lines

doi: 10.1016/j.bbrc.2020.03.061

Figure Lengend Snippet: The co-culture of breast cancer cell lines with CD36+ fibroblasts showed growth inhibition, reversion of basal and lateral polarity, or reduced malignant potential. (a-d) Co-culture of MDA-MB-231 with CD36+ or CD36− fibroblasts resulted in growth inhibition or promotion, respectively; two populations of Ki67 expressing cells; and higher frequency of proliferating cells, with high-level of Ki67, with the CD36− co-culture. (e-g) Co-culture of MCF7 with CD36+ induced renormalization of lateral and basal polarity, reduced colony formation, and reduced number of single cells. (h-i) Preconditioned MDA-MB-231 cells to CD36+ fibroblasts show reduced malignant potential as indicated with the AIG assay, and preconditioned MCF7 cells to CD36+ fibroblasts show no significant statistical difference.

Article Snippet: CD36 transfection and validation Fibroblasts were transfected with CD36 overexpression and knockdown constructs using CRISPR-cas9 technology (Santa Cruz Biotechnology SC400233-ACT/KO) and validated using fluorescent microscopy per vendor’s protocol.

Techniques: Co-Culture Assay, Inhibition, Expressing

Journal: Biochemical and biophysical research communications

Article Title: Overexpression of CD36 in mammary fibroblasts suppresses colony growth in breast cancer cell lines

doi: 10.1016/j.bbrc.2020.03.061

Figure Lengend Snippet: YY1 and pSmad 2/3 expressions were downregulated in organoid models of MCF7 and MDA-MB-231, co-cultured with CD36+ fibroblasts, respectively.

Article Snippet: CD36 transfection and validation Fibroblasts were transfected with CD36 overexpression and knockdown constructs using CRISPR-cas9 technology (Santa Cruz Biotechnology SC400233-ACT/KO) and validated using fluorescent microscopy per vendor’s protocol.

Techniques: Cell Culture