Journal: Scientific Reports

Article Title: The BET inhibitor attenuates the inflammatory response and cell migration in human microglial HMC3 cell line

doi: 10.1038/s41598-021-87828-1

Figure Lengend Snippet: Effects of LPS and JQ1 on HMC3 cells and the expression of inflammatory response-related genes in LPS- and JQ1-treated cells. ( A ) Morphology of HMC3 cells after treatment with LPS, JQ1, and LPS + JQ1 for 4 h. ( B ) HMC3 cells were cultured for 4 h after treatment with LPS (100 ng/ml) and JQ1 (500 nM) and stained for IBA1 (green fluorescence), a microglia-lineage marker. Nuclei were counterstained with DAPI (blue). ( C ) HMC3 cells were treated with LPS and JQ1 at different concentrations for different durations (4 h, 24 h, and 48 h). The viability of the HMC3 cells was determined using the WST-1 assay. ( D ) HMC3 cells were treated with LPS at different doses (10, 100, and 1000 ng/ml) for 4 h. Inflammatory genes were significantly upregulated in cells treated with LPS compared to DMSO-treated control cells (upper panel). HMC3 cells were treated with LPS (100 ng/ml) and JQ1 at different doses (50, 500, and 5000 nM) for 4 h. Inflammatory genes were significantly downregulated in cells treated with JQ1 (bottom panel). Gene expression was normalized to GAPDH transcript levels. The data represent three independent experiments. The values are the mean ± SEM of triplicate experiments (* p < 0.05 and ** p < 0.001).

Article Snippet: HMC3 human microglial cells were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Expressing, Cell Culture, Staining, Fluorescence, Marker, WST-1 Assay, Control, Gene Expression

Journal: Scientific Reports

Article Title: The BET inhibitor attenuates the inflammatory response and cell migration in human microglial HMC3 cell line

doi: 10.1038/s41598-021-87828-1

Figure Lengend Snippet: Differentially expressed genes in LPS- and LPS + JQ1-treated HMC3 cells. ( A ) Heat map showing the top 50 upregulated genes in HMC3 cells treated with LPS and LPS + JQ1 for 4 h determined by RNA-seq ( p ≤ 0.05 and fold change ≥ 1.5 log2). Each experiment was performed in experimental triplicates (n = 3) for each condition, and the results were individually combined. The color scale shown in the heat map represents the log 2 fold change values. Red cells indicate upregulated gene, while blue cells indicate downregulated genes. The p-value with an asterisk attached in the cell represents * p < 0.05, ** p < 0.01, and *** p < 0.001. ( B ) GO term analysis of the biological processes associated with up- and downregulated genes after LPS (100 ng/ml) and JQ1 (500 nM) treatment for 4 h. GO analysis of the number of genes shown in the van diagram. ( C ) Biological pathway analysis of DEGs in cells treated with LPS and LPS + JQ1 using IPA (QIAGEN Inc., https://www.qiagenbioinformatics.com/products/ingenuitypathway-analysis ) showed up- and downregulated genes.

Article Snippet: HMC3 human microglial cells were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: RNA Sequencing

Journal: Scientific Reports

Article Title: The BET inhibitor attenuates the inflammatory response and cell migration in human microglial HMC3 cell line

doi: 10.1038/s41598-021-87828-1

Figure Lengend Snippet: JQ1 inhibits a specific subset of LPS-inducible inflammatory genes. ( A ) Heat map showing chemokine/cytokine and interferon response gene expression in HMC3 cells treated with LPS (100 ng/ml) and JQ1 (500 nM) for 4 h. The color scale shown in the heat map represents the log 2 fold change values. ( B ) IPA downstream effects analysis presenting the mapping of most significant genes related to inflammatory response genes in LPS-treated cells. The DEGs are colored by predicted activation state, such as activated (red) or suppressed (green). The edges connecting the nodes are colored orange (leading to activation of the downstream node), blue (leading to inhibition), and yellow (if the findings underlying the relationship are inconsistent with the state of the downstream node). ( C ) Expression levels of inflammation-related genes were analyzed by qRT-PCR and normalized to GAPDH transcript levels. Overall, LPS-induced inflammatory genes were inhibited by JQ1. The data represent three independent experiments. The values are the mean ± SEM of triplicate experiments (** p < 0.001).

Article Snippet: HMC3 human microglial cells were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Gene Expression, Activation Assay, Inhibition, Expressing, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: The BET inhibitor attenuates the inflammatory response and cell migration in human microglial HMC3 cell line

doi: 10.1038/s41598-021-87828-1

Figure Lengend Snippet: IRF1 regulates migration-related genes in LPS- and LPS + JQ1-treated HMC3 cells. ( A ) IPA upstream regulator analysis of DEGs in LPS-, JQ1- and LPS + JQ1-treated HMC3 cells. The color scale shown in the heat map represents the activation z-score value. ( B ) TF-binding motif analysis to determine TF-binding site enrichment in the promoters of migratory genes. Putative binding sites for IRF1was significantly enriched in LPS-induced migratory genes. ( C ) Expression of RELA and IRF family genes were analyzed by qRT-PCR and normalized to GAPDH transcript levels. Among these IRF family genes, IRF1 and IRF2 were significantly upregulated in LPS-treated cells. The data represent three independent experiments. The values are the mean ± SEM of triplicate experiments (** p < 0.001). ( D ) IPA upstream regulator analysis identified key regulatory gene IRF1, and it targets genes in LPS-treated cells. Of the genes regulated by IRF1, only migration-related genes are shown in bold and in red. The migration-related genes were highly correlated with IRF1. ( E ) Log 2 fold changes and p -value in the expression of IRF1 target genes. ( F ) IPA network analysis of genes related to the inflammatory response and migration in LPS-treated cells. Of the genes identified, only chemotaxis-related genes are shown in bold and in red. Most inflammation-related genes overlap with migration-related genes.

Article Snippet: HMC3 human microglial cells were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Migration, Activation Assay, Binding Assay, Expressing, Quantitative RT-PCR, Chemotaxis Assay

Journal: Physiological Research

Article Title: Berberine Exerts Neuroprotective Effects in Alzheimer’s Disease by Switching Microglia M1/M2 Polarization Through PI3K-AKT Signaling

doi: 10.33549/physiolres.935410

Figure Lengend Snippet: ( A ) CCK8 assay was conducted to analyze the toxicity of different doses of BBR (1 μM, 2 μM, 4 μM, 8 μM, or 16 μM) in HMC3 cells. ( B-F ) HMC3 cells were divided into six groups: control, BBR (8 μM), LPS, LPS+BBR (2 μM), LPS+BBR (4 μM), and LPS+BBR (8 μM). For BBR and LPS co-treated group, HMC3 cells were treated with BBR for 3 h, followed by LPS exposure for 24 h. ( B and C ) IF assay was carried out to evaluate the levels of two indicators of inflammation, including iNOS and COX2. ( D-F ) ELISA was implemented to analyze the release of three pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in the culture supernatant. ns: no statistically significant difference, * P< 0.05, ** P< 0.01, *** P< 0.001.

Article Snippet: Human microglia cell line HMC3 and human neuroblastoma cell line SH-SY5Y were purchased from Procell Biotechnology (Wuhan, China).

Techniques: CCK-8 Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Physiological Research

Article Title: Berberine Exerts Neuroprotective Effects in Alzheimer’s Disease by Switching Microglia M1/M2 Polarization Through PI3K-AKT Signaling

doi: 10.33549/physiolres.935410

Figure Lengend Snippet: ( A-D ) Aβ 1-42 -exposed SH-SY5Y cells were cultured in the conditioned medium from control, BBR-stimulated, LPS-stimulated, or BBR and LPS co-treated HMC3 cells. ( A ) CCK8 assay was employed to analyze the viability of SH-SY5Y cells. ( B and C ) The MMP and ROS in SH-SY5Y cells were determined through IF staining method of JC-1 and DCFH-DA, respectively. ( D ) Western blot assay was conducted to measure the protein levels of two apoptosis-associated markers (cleaved CASP3 and Bax) in SH-SY5Y cells. ns: no statistically significant difference, * P< 0.05, *** P< 0.001.

Article Snippet: Human microglia cell line HMC3 and human neuroblastoma cell line SH-SY5Y were purchased from Procell Biotechnology (Wuhan, China).

Techniques: Cell Culture, Control, CCK-8 Assay, Staining, Western Blot

Journal: Physiological Research

Article Title: Berberine Exerts Neuroprotective Effects in Alzheimer’s Disease by Switching Microglia M1/M2 Polarization Through PI3K-AKT Signaling

doi: 10.33549/physiolres.935410

Figure Lengend Snippet: ( A-C ) HMC3 cells were divided into five groups: control, BBR (8 μM), LPS, LPS+BBR (8 μM), LPS+BBR (8 μM) + LY294002. For BBR and LPS co-treated group, HMC3 cells were treated with BBR for 3 h, followed by LPS exposure for 24 h. For LPS, BBR, and LY294002 co-treated group, HMC3 cells were treated with BBR for 3 h, followed by LPS and LY294002 stimulation for 24 h. ( A ) The levels of p-PI3K, PI3K, p-AKT, and AKT in HMC3 cells were determined by western blot assay. ( B and C ) ELISA was employed to detect the release of two pro-inflammatory cytokines (IL-1β and IL-6) in the culture supernatant of HMC3 cells. ns: no statistically significant difference, ** P< 0.01, *** P< 0.001.

Article Snippet: Human microglia cell line HMC3 and human neuroblastoma cell line SH-SY5Y were purchased from Procell Biotechnology (Wuhan, China).

Techniques: Control, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Bioengineered

Article Title: LINC01088 promotes the growth and invasion of glioma cells through regulating small nuclear ribonucleoprotein polypeptide A transcription

doi: 10.1080/21655979.2022.2051786

Figure Lengend Snippet: The expression profile, cellular location, and distribution of LINC01088 . (a) GEPIA ( http://gepia.cancer-pku.cn/index.html ) results of up-expression of LINC01088 in glioblastoma multiforme (GBM) and brain lower-grade glioma (LGG) tissues. (b) The volcano map of differentially expressed lncRNAs in GSE15824. (c) qRT-PCR analysis of LINC01088 expression in microglia cell line HMC3 and glioma cell lines (U251, T98G, SHG-44 and U87). (d) The expression and location of LINC01088 (green) in U251 and T98G cells was determined by FISH assay with confocal microscopy. DAPI-stained nuclei were blue. Scar bar: 50 μM. (e) The expression level of LINC01088 in the subcellular fractions of U251 and T98G cells was detected by qRT-PCR. All data were presented as means ± SD of three independent experiments. Values are significant at * P < 0.5 and ** P < 0.01 as indicated.

Article Snippet: A human microglia cell line HMC3 and glioma cell line U251 and T98G were purchased from Procell Life Science&Technology Co., Ltd (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Confocal Microscopy, Staining

Journal: Theranostics

Article Title: KLK8/HGF/Met signaling pathway mediates diabetes-associated hippocampal neuroinflammation in male mice

doi: 10.7150/thno.109513

Figure Lengend Snippet: KLK8 promotes microglial activation via a Met-dependent signaling pathway. A-B , BV2 mouse microglial cells were infected with Ad-KLK8 at a MOI of 1, 3, or 10 for 48 h. A , mRNA and protein expression levels of Iba1 were detected by qRT-PCR (F 3,12 = 167.971, p < 0.001) and western blotting (F 3,12 = 124.428, p < 0.001), respectively. Representative protein bands were presented on the left of the histograms. B , The mRNA expression levels of TNF-α, IL-6, CCL2, and iNOS were detected by qRT-PCR (TNF-α: F 3,12 = 225.322, p < 0.001. IL-6: F 3,12 = 23.992, p < 0.001. CCL2: F 3,12 = 59.73, p < 0.001. iNOS: F 3,12 = 127.568, p < 0.001). C-E , BV2 cells were infected with Ad-Vector or Ad-KLK8 at a MOI of 3 for 48 h. Dysregulated genes were analyzed by RNA-seq. C , Volcano plots showing DEGs in Ad-KLK8-treated BV2 cells. Red reflects upregulated and blue indicates downregulated genes. D, Heat map of DEGs, red indicates upregulated and blue shows downregulated genes. E , GSEA was then performed to determine the enriched signaling pathways in KLK8-overexpressed BV2 cells. GSEA gene sets associated with Met-activates-PTK2 (top) and Met-promotes-cell-motility (bottom) signaling pathways were significantly enriched in the Ad-KLK8-treated BV2 cells. Heat maps of the dysregulated target genes of Met-activates-PTK2 and Met-promotes-cell-motility (bottom) signaling pathways in Ad-KLK8 treated BV2 were presented on the right of the GSEA plots. Red reflects upregulated and blue indicates downregulated genes. F-G , BV2 cells were infected with Ad-KLK8 at a MOI of 3 for 48 h in the presence or absence of the Met inhibitor JNJ-38877605 at the indicated concentrations. H-I, A stably KLK8-overexpressing HMC3 cell line was generated through infection with Lv-KLK8. Cells infected with an empty lentivirus served as the control group (Lv-Vector). Stably KLK8-overexpressing HMC3 cells and control cells were treated with or without the Met inhibitor JNJ-38877605 at the indicated concentrations for 48 h. Protein levels of p-Met, Met, p-Btk, Btk, p-p65, p65, Src, and Iba1 in BV2 cells (F) or HMC3 cells (H) were determined by western blot analysis . The mRNA expression levels of Iba1, TNF-α, IL-6, CCL2, and iNOS in BV2 cells (G, Iba1: F 5,18 = 68.758, p < 0.001. TNF-α: F 5,18 = 43.501, p < 0.001. IL-6: F 5,18 = 259.947, p < 0.001. CCL2: F 5,18 = 61.644, p < 0.001. iNOS: F 5,18 = 111.889, p < 0.001) or HMC3 cells (I, Iba1: F 5,18 = 23.051, p < 0.001. TNF-α: F 5,18 = 17.202, p < 0.001. IL-6: F 5,18 = 61.04, p < 0.001. CCL2: F 5,18 = 12.358, p < 0.001. iNOS: F 5,18 = 19.215, p < 0.001 ) were detected by qRT-PCR . Data were presented as means ± SEM (n = 4, one-way ANOVA). * p < 0.05, ** p < 0.01. JNJ represents JNJ-38877605.

Article Snippet: HMC3 human microglial cell lines were obtained from Servicebio (Wuhan, China) and cultivated in DMEM (Gibco, New York, USA) containing 10% FBS (Gibco, New York, USA), and 1% penicillin/streptomycin (Gibco, New York, USA) at 37 °C in a humidified atmosphere consisting of 95% O 2 and 5% CO 2 .

Techniques: Activation Assay, Infection, Expressing, Quantitative RT-PCR, Western Blot, Plasmid Preparation, RNA Sequencing, Protein-Protein interactions, Stable Transfection, Generated, Control

Journal: Theranostics

Article Title: KLK8/HGF/Met signaling pathway mediates diabetes-associated hippocampal neuroinflammation in male mice

doi: 10.7150/thno.109513

Figure Lengend Snippet: KLK8 cleaves pro-HGF and augments HGF release, thereby facilitating Met signaling and microglial activation. A, BV2 cells were infected with Ad-KLK8 at a MOI of 3 for 48 h with or without anti-KLK8 neutralizing antibody (2.5 μg/mL). HGF contents in the cell medium were measured by ELISA assay (F 3,12 = 16.994, p < 0.001). B, BV2 cells were infected with Ad-KLK8 at a MOI of 3 for 48 h with or without serine protease inhibitors ZnSO4 (0.05 mM) and antipain (0.05 mM), respectively (left, F 5,18 = 39.434, p < 0.001). A stably KLK8-overexpressing HMC3 cell line was generated through infection with Lv-KLK8. Cells infected with an empty lentivirus served as the Lv-Vector. Stably KLK8-overexpressing HMC3 cells and control cells were treated with or without serine protease inhibitors ZnSO4 (0.05 mM) and antipain (0.05 mM), respectively (right, F 5,18 = 14.914, p < 0.001). HGF contents in the cell medium were measured by ELISA assay. C , Purified recombinant human pro-HGF (rhpro-HGF) was incubated with or without activated recombinant human KLK8 (rhKLK8) at the indicated concentrations, and analyzed by western blot using a pro-HGF antibody. D-E , A stably KLK8-overexpressing HMC3 cell line was generated through infection with Lv-KLK8. Cells infected with an empty lentivirus served as the Lv-Vector. Stably KLK8-overexpressing HMC3 cells and control cells were treated with or without a fully human anti-HGF neutralizing antibody Rilotumumab at the indicated concentrations. D , Protein levels of p-Met, Met, p-Btk, Btk, p-p65, p65, Src, and Iba1 were determined by western blot analysis. Representative protein bands were presented on the left of the histograms (p-Met/Met: F 3,12 = 50.48, p < 0.001. p-Btk/Btk: F 3,12 = 90.277, p < 0.001. p-p65/p65: F 3,12 = 89.791, p < 0.001. Src/β-actin: F 3,12 = 555.454, p < 0.001. Iba1/β-actin: F 3,12 = 141.437, p < 0.001). E, The mRNA levels of Iba1, TNF-α, IL-6, CCL2, and iNOS were detected by qRT-PCR ( Iba1: F 5,18 = 44.23, p < 0.001. TNF-α: F 5,18 = 52.255, p < 0.001. IL-6: F 5,18 = 99.978, p < 0.001. CCL2: F 5,18 = 41.607, p < 0.001. iNOS: F 5,18 = 77.973, p < 0.001). F , Schematic diagram of the mechanism underlying KLK8-induced microglial activation. KLK8 cleaves pro-HGF and augments HGF release, leading to the activation of the Met/Src/BTK/NF-κB signaling pathway and subsequent microglial activation. Data were presented as means ± SEM (n = 4, one-way ANOVA). * p < 0.05, ** p < 0.01.

Article Snippet: HMC3 human microglial cell lines were obtained from Servicebio (Wuhan, China) and cultivated in DMEM (Gibco, New York, USA) containing 10% FBS (Gibco, New York, USA), and 1% penicillin/streptomycin (Gibco, New York, USA) at 37 °C in a humidified atmosphere consisting of 95% O 2 and 5% CO 2 .

Techniques: Activation Assay, Infection, Enzyme-linked Immunosorbent Assay, Stable Transfection, Generated, Plasmid Preparation, Control, Purification, Recombinant, Incubation, Western Blot, Quantitative RT-PCR

Journal: Chemical Science

Article Title: Monitoring phagocytic uptake of amyloid β into glial cell lysosomes in real time †

doi: 10.1039/d1sc03486c

Figure Lengend Snippet: Synthesis and characterization of Aβ pH . (A) The Aβ pH is synthesized by conjugating the amine-reactive pH-sensitive Protonex Green dye to the side chain amine groups of the lysine residues and the N-terminal of human Aβ 1–42 peptide. (B) The pH-sensitivity of the Aβ pH probe characterized at different concentrations from 0.1 μM to 5.0 μM. Increased fluorescence is observed at acidic pH values of ∼5.0 to ∼2.0, covering the pH range of the intracellular acidic organelles. (C) Atomic force microscopy topographic images of Aβ pH oligomers compared to synthetic Aβ oligomers. Left-2D topographic image of Aβ pH and synthetic Aβ oligomers. Right-3D image (2 × 2 μm x – y ). (D) Live cell imaging of the phagocytic uptake of 1 μM Aβ pH by BV2 and N9 mouse microglia and by HMC3 human microglia over 24 hours. (E) Quantification of Aβ pH phagocytic score by BV2, N9, and HMC3 microglial cells from the live cell images. (F) The phagocytic uptake of Aβ pH by BV2 cells is measured and quantified via flow cytometry analysis. Dot plot shows live (PI − ) and Aβ pH+ cells. No green fluorescence is measured in unstained cells (UC) and in dead cells stained with the PI only whereas green fluorescence is measured in cells treated with 0.5 and 5.0 μM Aβ pH for 1 hour (higher fluorescence is seen in cells exposed to the higher concentration of Aβ pH ). Data shown in terms of % max, by scaling each curve to mode = 100% ( y -axis).

Article Snippet: HMC3 human microglial cell line was a gift from Dr Jianming Li (Purdue University, USA) who originally obtained the cells from ATCC.

Techniques: Synthesized, Fluorescence, Microscopy, Live Cell Imaging, Flow Cytometry, Staining, Concentration Assay

Journal: Chemical Science

Article Title: Monitoring phagocytic uptake of amyloid β into glial cell lysosomes in real time †

doi: 10.1039/d1sc03486c

Figure Lengend Snippet: Fluorescence of internalized Aβ pH is retained in fixed cells. (A) Confocal images of fixed HMC3, N9, and BV2 cells showing the uptake of Aβ pH (green). Cells are stained for acidic intracellular organelles (LysoTracker Red, confirming co-localization of the Aβ pH within the acidic intracellular organelles) and nuclei (DAPI, blue). No antibody is required to detect Aβ pH . (B) Primary mouse microglia grown in defined, reduced-serum media phagocytose Aβ pH ex vivo . Cells are fixed and stained for nuclei and show Aβ pH colocalized in the acidic organelles with LysoTracker Red. (C) The phagocytic uptake of Aβ pH by primary microglia is measured and quantified via flow cytometry analysis. Dot plot shows live (ZV − ) and Aβ pH+ cells. No green fluorescence is measured in unstained cells (UC) or dead cells stained with the ZV live/dead stain only whereas green fluorescence is measured in cells treated with 0.5, 1.0, and 2.0 μM Aβ pH for 1 hour. Data shown in terms of % max, by scaling each curve to mode = 100% ( y -axis). (D) Primary immunopanned rat astrocytes also phagocytose Aβ pH in serum-free conditions. Cells are fixed and stained for astrocyte specific GFAP antibody (red) and nuclei. (E) Uptake of Aβ pH over time by primary immunopanned astrocytes as observed in live cells in real time. (F) Quantification of uptake of 0.5, 1.0, and 2.0 μM Aβ pH by primary astrocytes. Data are mean ± SEM, n = 8 separate wells per group/timepoint.

Article Snippet: HMC3 human microglial cell line was a gift from Dr Jianming Li (Purdue University, USA) who originally obtained the cells from ATCC.

Techniques: Fluorescence, Staining, Ex Vivo, Flow Cytometry

Journal: Actas Españolas de Psiquiatría

Article Title: Quercetin Regulates the Polarization of Microglia through the NRF2/HO1 Pathway and Mitigates Alzheimer's Disease

doi: 10.62641/aep.v52i6.1713

Figure Lengend Snippet: Que increases cell viability and NRF2/HO1 pathway activation in microglial cells . (A,B) Cell counting kit-8 (CCK-8) assay results showing cell viability of human microglial cells HMC3 treated with different concentrations of Que (A), and with A β (1-42) and different concentrations of Que (B) (ANOVA). (C) Levels of NRF2/HO1 pathway-related proteins (ANOVA). n = 5. Control, HMC3 cells with no treatments; A β (1-42), HMC3 cells treated with A β (1-42) (1 µM); A β (1-42) + Que, HMC3 cells treated with A β (1-42) (1 µM) and Que (5 µM); A β (1-42) + Que + ML385, HMC3 cells treated with A β (1-42) (1 µM), Que (5 µM), and the NEF2 selective inhibitor ML385 (5 µM). * p < 0.05, ** p < 0.01, *** p < 0.001. Abbreviations: Que, quercetin; A β (1-42), amyloid-beta 1-42; ML385, NRF2 inhibitor; NRF2, nuclear factor E2-related factor 2; Histone-3, Histone H3; HO1, heme oxygenase-1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Human microglial cells HMC3 (iCell-h301, iCell Biosciences Inc., Shanghai, China) were cultured in a specific medium (iCell-h301-001b, iCell Biosciences Inc., Shanghai, China) at 37 °C, 70%–80% humidity, and 5% CO 2 .

Techniques: Activation Assay, Cell Counting, CCK-8 Assay, Control

Journal: Actas Españolas de Psiquiatría

Article Title: Quercetin Regulates the Polarization of Microglia through the NRF2/HO1 Pathway and Mitigates Alzheimer's Disease

doi: 10.62641/aep.v52i6.1713

Figure Lengend Snippet: Que activates the NRF2/HO1 pathway in microglial cells . (A) Lactate dehydrogenase (LDH) release levels in HMC3 cells measured by enzyme-linked immunosorbent assay (ELISA) (ANOVA). (B,C) Quantification (B) and representative images (C; scale bar: 50 µm) of apoptotic cells (ANOVA). n = 5. *** p < 0.001. Abbreviations: Que, quercetin; A β (1-42), amyloid-beta 1-42; ML385, NRF2 inhibitor; LDH, lactate dehydrogenase; TUNEL, terminal deoxynucleotidyl transferase dUTP nick-end labeling; DAPI, 4 ′ ,6-diamidino-2-phenylindole.

Article Snippet: Human microglial cells HMC3 (iCell-h301, iCell Biosciences Inc., Shanghai, China) were cultured in a specific medium (iCell-h301-001b, iCell Biosciences Inc., Shanghai, China) at 37 °C, 70%–80% humidity, and 5% CO 2 .

Techniques: Enzyme-linked Immunosorbent Assay, TUNEL Assay

Journal: Actas Españolas de Psiquiatría

Article Title: Quercetin Regulates the Polarization of Microglia through the NRF2/HO1 Pathway and Mitigates Alzheimer's Disease

doi: 10.62641/aep.v52i6.1713

Figure Lengend Snippet: Que decreases neuroinflammation and oxidative stress in HMC3 cells by activating the NRF2/HO1 pathway . (A) mRNA levels of inflammation cytokines (ANOVA). (B–D) Protein levels of inflammation cytokines detected by ELISA (ANOVA). (E–G) Protein levels of oxidative stress-related markers: malondialdehyde (MDA; E), superoxide dismutase (SOD; F), and reduced glutathione (GSH)/oxidized glutathione disulfide (GSSG; G) detected by ELISA (ANOVA). n = 5. ** p < 0.01, *** p < 0.001. Abbreviations: A β (1-42), amyloid-beta 1-42; Que, quercetin; ML385, NRF2 inhibitor; IL, interleukin; TNF- α , tumor necrosis factor alpha; MDA, malondialdehyde; SOD, superoxide dismutase; GSH/GSSG, glutathione/oxidized glutathione disulfide.

Article Snippet: Human microglial cells HMC3 (iCell-h301, iCell Biosciences Inc., Shanghai, China) were cultured in a specific medium (iCell-h301-001b, iCell Biosciences Inc., Shanghai, China) at 37 °C, 70%–80% humidity, and 5% CO 2 .

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Cellular and Molecular Neurobiology

Article Title: Proinflammatory Extracellular Vesicle-Mediated Signaling Contributes to the Induction of Neuroinflammation in Animal Models of Endotoxemia and Peripheral Surgical Stress

doi: 10.1007/s10571-020-00905-3

Figure Lengend Snippet: Effect of LPS- or HpX-derived EVs on cultured glia cells. EVs from LPS- ( a ) or HpX- ( b ) treated animals were added to the culture medium of HMC3 glia cells at the indicated concentrations. After 1 h the conditioned medium was assayed for TNF-α, IL1ß and IL6. Results are the means of 6 independent observations. Error bars indicate S.D. #Below detection limit of the assay. *** p < 0.001. In the case that values were below detection limit of the assay, no statistical analysis could be performed

Article Snippet: Human microglial HMC3 cells (Dello Russo et al. ) were obtained from LGC Standards GmbH (Wesel, Germany) and cultured in 96 well plates with DMEM medium containing 10% fetal calf serum (Sigma-Aldrich, Taufkirchen, Germany) and 1% penicillin/streptomycin.

Techniques: Derivative Assay, Cell Culture