Journal: Journal of Advanced Research

Article Title: Kaempferol regulating macrophage foaming and atherosclerosis through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway

doi: 10.1016/j.jare.2024.11.016

Figure Lengend Snippet: KAE inhibited ox-LDL-induced inflammation in RAW264.7 macrophage. (A) Representative images of Oil red “O” staining in RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL. Scale bar: 50 μm. (B) Statistical results of Oil red “O” staining from (A) (n = 5). (C-F) The mRNA levels of IL1b, IL6, TNFα, and MCP1 by RT-PCR in RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL (n = 5). (G-J) The protein expression of IL-6, TNF-α, IL-1β, and MCP-1 by ELISA in RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL (n = 5). The results represent means ± SEM. *P < 0.05, **P < 0.01 was compared with the control group. # P < 0.05, ## P < 0.01 was compared with the ox-LDL group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: RAW264.7 cells (ATCC, USA) were cultured in DMEM containing 10 % fetal bovine serum (FBS, Gibco, USA) and 1 % penicillin/streptomycin solution (P/S) and stored at 37°C in a 5 % CO 2 incubator.

Techniques: Staining, Reverse Transcription Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Advanced Research

Article Title: Kaempferol regulating macrophage foaming and atherosclerosis through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway

doi: 10.1016/j.jare.2024.11.016

Figure Lengend Snippet: KAE inhibited ox-LDL-induced CD36 expression and Ca 2+ influx. (A) Histogram showing CD36 + macrophages in RAW264.7 macrophage after 100 µg/mL ox-LDL treatment were quantified by FCM. Statistical results of the proportion (B) and MFI (C) from (A) (n = 3). (D-E) The protein levels of CD36 by Western Blot in RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL and KAE(n = 3). (F) The fluorescence images of CD36 in different groups of RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL. (G) Statistical results of fluorescence images from (F) (n = 4). (H) Representative fluorescence images of Ca 2+ in different groups of RAW264.7 macrophage (I) Statistical results of fluorescence images from (H) (n = 4). The results represent means ± SEM. *P < 0.05, **P < 0.01 was compared with the control group. # P < 0.05 was compared with the ox-LDL group.

Article Snippet: RAW264.7 cells (ATCC, USA) were cultured in DMEM containing 10 % fetal bovine serum (FBS, Gibco, USA) and 1 % penicillin/streptomycin solution (P/S) and stored at 37°C in a 5 % CO 2 incubator.

Techniques: Expressing, Western Blot, Fluorescence, Control

Journal: Journal of Advanced Research

Article Title: Kaempferol regulating macrophage foaming and atherosclerosis through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway

doi: 10.1016/j.jare.2024.11.016

Figure Lengend Snippet: KAE inhibits ox-LDL-induced mitochondrial oxidative damage in RAW264.7 cells. (A) The images of DCFH-DA staining in RAW264.7 cells after treatment with 100 µg/mL ox-LDL. Scale bar: 50 μm. (B) The images of JC-10 staining in RAW264.7 macrophages after treatment with 100 µg/mL ox-LDL. Scale bar: 50 μm. (C) Statistical results of JC-10 staining from (B) (n = 4). The results represent means ± SEM. **P < 0.01 was compared with the control group. ## P < 0.01 was compared with the ox-LDL group.

Article Snippet: RAW264.7 cells (ATCC, USA) were cultured in DMEM containing 10 % fetal bovine serum (FBS, Gibco, USA) and 1 % penicillin/streptomycin solution (P/S) and stored at 37°C in a 5 % CO 2 incubator.

Techniques: Staining, Control

Journal: Journal of Advanced Research

Article Title: Kaempferol regulating macrophage foaming and atherosclerosis through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway

doi: 10.1016/j.jare.2024.11.016

Figure Lengend Snippet: KAE inhibits inflammatory response through MAPK/NF-κB and Nrf2/HO-1 pathways. (A-D) The protein levels of p-JNK, JNK, p-ERK, ERK, p-p38 and p38 by Western Blot in RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL and kaempferol (n = 3). (E-F) The protein expression of p65 and p-p65 by Western Blot in RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL and kaempferol (n = 3). (G-I) The protein levels of, Nrf2 and HO-1 by Western Blot in RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL (n = 3). (J) Representative fluorescence images of Nrf2 in different groups of RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL. (K) Statistical results of fluorescence images from (J) (n = 4). The results represent means ± SEM. *P < 0.05, **P < 0.01 was compared with the control group. # P < 0.05 was compared with the ox-LDL group.

Article Snippet: RAW264.7 cells (ATCC, USA) were cultured in DMEM containing 10 % fetal bovine serum (FBS, Gibco, USA) and 1 % penicillin/streptomycin solution (P/S) and stored at 37°C in a 5 % CO 2 incubator.

Techniques: Western Blot, Expressing, Fluorescence, Control

Journal: Journal of Advanced Research

Article Title: Kaempferol regulating macrophage foaming and atherosclerosis through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway

doi: 10.1016/j.jare.2024.11.016

Figure Lengend Snippet: KAE suppresses the expression of Piezo1 in ApoE -/- mice and Yoda1 induced Ca 2+ influx in RAW264.7 macrophages. (A) The images of immunofluorescence staining with Piezo1 (red) and CD68 (green) in aortic root regions. Scale bar: 50 μm. (B) Typical trace of 5 μM Yoda1-induced intracellular Ca 2+ response with/without kaempferol in RAW264.7 cells (N = 8 each). (C) Statistical data from (B) (n = 3). (D) Representative fluorescence images of Ca 2+ of RAW264.7 cells after treatment with 5 μM Yoda1. (E) Statistical results of fluorescence images from (D) (n = 4). The results represent means ± SEM. *P < 0.05, **P < 0.01 was compared with the control group. # P < 0.05 was compared with the model group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: RAW264.7 cells (ATCC, USA) were cultured in DMEM containing 10 % fetal bovine serum (FBS, Gibco, USA) and 1 % penicillin/streptomycin solution (P/S) and stored at 37°C in a 5 % CO 2 incubator.

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Control

Journal: NPJ Biofilms and Microbiomes

Article Title: In silico identification of two peptides with antibacterial activity against multidrug-resistant Staphylococcus aureus

doi: 10.1038/s41522-022-00320-0

Figure Lengend Snippet: Inhibitory action of HG2 and HG4 on LPS or LTA-driven inflammation in murine macrophages: eLUCidate™ Raw 264.7 NF-kB reporter cell line was used to measure the inhibitory action of HG2 and HG4 on LPS ( a ) or LTA ( b ) -mediated inflammation as explained in the Materials and Methods’ section. Data were plotted using GraphPad® Prism 7 software. Results are expressed as means ± standard deviation. Representative images of toxicity assay of peptides ( c ) — ( i ) HG2 and ( ii ) HG4 in G. mellonella 120 h post-treatment with 3x MIC concentrations. The larvae remained alive and without melanisation. Virulence assay of MRSA USA300 in G. mellonella using a lethal dose inoculum of 10 6 CFU/per larvae—( iii ) 24 h post-infection: some larvae were dead and partial melanisation was observed. ( iv ). 48 h post-infection: most larvae were dead and complete melanisation was evident. The experiment was done with three experimental replicates, each containing groups of 10 larvae. d Kaplan–Meier survival curves of G. mellonella infected with a lethal dose (LD 50 ) of S. aureus (2.25 × 10 6 CFU/larvae) and treated with peptides HG2 and HG4 at 1x and 3x MIC concentrations and uninfected larvae treated with peptides in PBS at 3x MIC (placebo showing a 100% larvae survival rate).

Article Snippet: The inhibitory activity of HG2, HG4 and comparator compounds against LPS and LTA was assessed using eLUCidate™ Raw 264.7 NF-kB reporter cell line (AMSBIO) as previously described .

Techniques: Software, Standard Deviation, Infection

Journal: Microorganisms

Article Title: A Plasmid-Encoded FetMP-Fls Iron Uptake System Confers Selective Advantages to Salmonella enterica Serovar Typhimurium in Growth under Iron-Restricted Conditions and for Infection of Mammalian Host Cells

doi: 10.3390/microorganisms8050630

Figure Lengend Snippet: Invasion of epithelial HeLa cells ( A ) and replication within RAW264.7 macrophages ( B ) by wild-type S. Typhimurium LSP 146/02, its Δ fetMP - flsDA isogenic derivative, and the latter strain complemented with the cloned fetMP - flsDA genes (LSP 146/02 Δ fetMP - flsDA C ). The data shown are the mean values ± standard deviations of three biological replicates. Statistical significance (**** p < 0.0001; *** p < 0.001; ** p < 0.01; * p < 0.05; n.s., not significant) was determined by one-way ANOVA and the Tukey´s post-test.

Article Snippet: HeLa and RAW264.7 cells (Cell Lines Service (CLS), Heidelberg, Germany) were propagated in a high-glucose (4.5 g/L) Dulbecco’s modified Eagle’s medium (DMEM) containing 4 mM glutamine (PAA, Cölbe, Germany) and 10% (HeLa) or 6% (RAW264.7) inactivated fetal calf serum (iFCS) (Sigma), at 37 °C in an atmosphere of 5% CO 2 .

Techniques: Clone Assay

Journal: Microorganisms

Article Title: A Plasmid-Encoded FetMP-Fls Iron Uptake System Confers Selective Advantages to Salmonella enterica Serovar Typhimurium in Growth under Iron-Restricted Conditions and for Infection of Mammalian Host Cells

doi: 10.3390/microorganisms8050630

Figure Lengend Snippet: Invasion of epithelial HeLa cells by S. Typhimurium ATCC 14028 harboring the cloned fetMP - flsDA genes (ATCC 14028 fetMP - flsDA T ) ( A ) and replication of the same strain within RAW264.7 macrophages ( B ). The data are the mean values ± standard deviations of three biological replicates. Statistical significance (**** p < 0.0001; *** p < 0.001; ** p < 0.01) was determined by one-way ANOVA and the Tukey´s post-test.

Article Snippet: HeLa and RAW264.7 cells (Cell Lines Service (CLS), Heidelberg, Germany) were propagated in a high-glucose (4.5 g/L) Dulbecco’s modified Eagle’s medium (DMEM) containing 4 mM glutamine (PAA, Cölbe, Germany) and 10% (HeLa) or 6% (RAW264.7) inactivated fetal calf serum (iFCS) (Sigma), at 37 °C in an atmosphere of 5% CO 2 .

Techniques: Clone Assay