human microglia hmc3 cells Search Results


crl 3304tm  (ATCC)
99
ATCC crl 3304tm
Crl 3304tm, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hmc3 cells  (MedChemExpress)
98
MedChemExpress hmc3 cells
Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of <t>HMC3</t> cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001
Hmc3 Cells, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+microglia+hmc3+cells/MPZL1%2C+Human/pm37085894-224-7-26
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human brain microglia cells hmc3  (ATCC)
96
ATCC human brain microglia cells hmc3
Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of <t>HMC3</t> cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001
Human Brain Microglia Cells Hmc3, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+microglia+hmc3+cells/HMC3%3B+Embryonic+Microglia+Clone+3%3B+Human/pm39348530-297-0-7
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hmc3 human microglial cell lines  (Purdue University Cytometry)
86
Purdue University Cytometry hmc3 human microglial cell lines
Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of <t>HMC3</t> cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001
Hmc3 Human Microglial Cell Lines, supplied by Purdue University Cytometry, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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microglia  (Celprogen Inc)
94
Celprogen Inc microglia
Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of <t>HMC3</t> cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001
Microglia, supplied by Celprogen Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+microglia+hmc3+cells/Human+Microglia+Primary+Cell+Culture/pm38397814-65-4-10
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microglial  (Nikon)
99
Nikon microglial
Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of <t>HMC3</t> cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001
Microglial, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+microglia+hmc3+cells/Objectives/pm34890698-138-8-20
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microglial cell line hmc3  (Procell Inc)
86
Procell Inc microglial cell line hmc3
Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of <t>HMC3</t> cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001
Microglial Cell Line Hmc3, supplied by Procell Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+microglia+hmc3+cells/cells+hmc3/pm41457142-40-2-7
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n a atcc n a hmc3 unspecified n a atcc n a  (ATCC)
99
ATCC n a atcc n a hmc3 unspecified n a atcc n a
Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of <t>HMC3</t> cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001
N A Atcc N A Hmc3 Unspecified N A Atcc N A, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+microglia+hmc3+cells/THP-1/pmc12406725__jciinsight-10-181903-s040-26-57-58
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n a atcc n a hmc3 unspecified n a atcc n a - by Bioz Stars, 2026-09
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f5 77 h  (ATCC)
90
ATCC f5 77 h
Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of <t>HMC3</t> cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001
F5 77 H, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human her2 breast cancer cells  (ATCC)
97
ATCC human her2 breast cancer cells
(A) Schematic outline of the cell culture. Three biological replicates were established for HMC3, HBEC5i and NHA. All cells were grown in DMEM-HG medium under serum-deprived conditions for 24 h after which the media was collected and pooled from the three brain cell types to treat the <t>SKBR3</t> cells for another 24 h. SKBR3-control cells were maintained in serum-free media for 48 h. (B) Phase contrast microscopy image of non-treated SKBR3 cells under 20X magnification. (C) Immunofluorescence microscopy of non-treated SKBR3 cells labeled with <t>anti-HER2</t> antibody and detected using an anti-mouse IgG secondary antibody conjugated to CruzFluor488. (D) Phase contrast microscopy image of CM-treated SKBR3 cells under 20X magnification shows development of filopodia/lamellipodia type protrusions from the surface of the cells. (E) Immunofluorescence microscopy of treated SKBR3 cells labeled with anti-HER2 antibody and detected using an anti-mouse IgG secondary antibody conjugated to CruzFluor488. (F) Stacked bar chart of percentage cells in the G1, S and G2 phases of the cell cycle for the control (C) and treated (T) SKBR3 cells.
Human Her2 Breast Cancer Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+microglia+hmc3+cells/SK-BR-3/bio_rxiv__2023__10__22__563488-41-0-19
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hmc3-mhciiluc:ubiaat gfp cells  (Neurix Inc)
90
Neurix Inc hmc3-mhciiluc:ubiaat gfp cells
(A) Schematic outline of the cell culture. Three biological replicates were established for HMC3, HBEC5i and NHA. All cells were grown in DMEM-HG medium under serum-deprived conditions for 24 h after which the media was collected and pooled from the three brain cell types to treat the <t>SKBR3</t> cells for another 24 h. SKBR3-control cells were maintained in serum-free media for 48 h. (B) Phase contrast microscopy image of non-treated SKBR3 cells under 20X magnification. (C) Immunofluorescence microscopy of non-treated SKBR3 cells labeled with <t>anti-HER2</t> antibody and detected using an anti-mouse IgG secondary antibody conjugated to CruzFluor488. (D) Phase contrast microscopy image of CM-treated SKBR3 cells under 20X magnification shows development of filopodia/lamellipodia type protrusions from the surface of the cells. (E) Immunofluorescence microscopy of treated SKBR3 cells labeled with anti-HER2 antibody and detected using an anti-mouse IgG secondary antibody conjugated to CruzFluor488. (F) Stacked bar chart of percentage cells in the G1, S and G2 phases of the cell cycle for the control (C) and treated (T) SKBR3 cells.
Hmc3 Mhciiluc:Ubiaat Gfp Cells, supplied by Neurix Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hcm3  (ATCC)
99
ATCC hcm3
A549 ( A ) and <t>HCM3</t> ( B ) cells were transfected for 24 h with pcDNA3 expressing rNS4B protein from ZIKV-15555 (rNS4B ZIKV-15555 ), MR766 (rNS4B MR766 ), or BeH819015 (rNS4B BeH819015 ). The rNS4B ZIK-15555 protein mutant lacking the 2K peptide is indicated as (Δ2K)-rNS4B ZIK-15555 . Cell lysates in RIPA buffer were assessed by immunoblot assays with anti-epitope antibody FLAG as the primary antibody. The β-tubulin protein served as a protein-loading control. The positions of rNS4B are indicated.
Hcm3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of HMC3 cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001

Journal: Genome biology

Article Title: YY1 lactylation in microglia promotes angiogenesis through transcription activation-mediated upregulation of FGF2.

doi: 10.1186/s13059-023-02931-y

Figure Lengend Snippet: Fig. 3 Hyperlactylation of microglia promotes angiogenesis in vitro. a Lactate production of HMC3 cells in the three groups treated with hypoxia for 0, 12, and 24 h (n = 3 independent experiments, at least 6 mice each group). b Quantification of Pan-Kla in the HMC3 cells of hypoxia 0 h, hypoxia 12 h, and hypoxia 24 h were analyzed by Western blotting (n = 3 per group). c Tube formation analysis. HMC3 cells were pretreated with 1% O2 or normoxia for 24 h and then cocultured with HRMECs; tube formation was assayed 12 h and 20 h after cell seeding (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. d HRMECs angiogenic capacity was evaluated by spheroid-sprouting assay. Representative images of spheroid sprouting were analyzed (n = 3 independent experiments, 3 images for each group); scale bar, 50 µm. e HRMECs migration was evaluated by Transwell assay (n = 3 independent experiments, 3 images for each group); scale bar, 25 µm. f HRMEC proliferation was evaluated by Ki67 staining (n = 3 independent experiments, 3 images for each group); scale bar, 100 µm. g, h Quantification of lactate and Pan-Kla in HMC3 cells in the three groups treated with DCA (20 mM), DMSO, and rotenone (50 nM) under hypoxia for 24 h (n = 3 per group). i–l HMC3 cells were pretreated with 1% O2 for 24 h with DCA, DMSO, or rotenone. Then HRMECs were cocultured with the pretreated microglia. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 100 μm (k) or 50 μm (i, j, and l). *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: RNA was isolated using TRIzol reagent from HMC3 cells (1 × 106 cells) or retinal tissue. cDNA was generated by the RT Master Mix for qPCR (MCE, HY-K0510).

Techniques: In Vitro, Western Blot, Migration, Transwell Assay, Staining

Fig. 4 The lactylation of YY1 in microglia plays an important role in regulating angiogenesis. a Heatmap shows the top 20 DELPs with increased and decreased lactylation under hypoxia for 24 h. b The enrichment analysis of DELPs. c Using the STRING database, DELPs specifically target the binding process. The node size corresponds to the relative fold-change of lactylation. d Collision-induced dissociation (CID) analysis was used to determine the modification sites. The MS/MS spectrum of modified “(Kla)SYLSGGA GAAGGGGADPGNK” is shown. e Hypoxia increases YY1 lactylation. Lactylation of YY1 in HMC3 cells under normoxia or hypoxia for 24 h was detected by YY1-Kla antibody (n = 3 per group). f–i HMC3 cells overexpressing WT or K183R YY1 were pretreated with 1% O2 for 24 h. Then HRMECs were cocultured with the pretreated HMC3 cells. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 50 μm (f, g, and i) or 100 μm (h). *p < 0.05; **p < 0.01; ***p < 0.001

Journal: Genome biology

Article Title: YY1 lactylation in microglia promotes angiogenesis through transcription activation-mediated upregulation of FGF2.

doi: 10.1186/s13059-023-02931-y

Figure Lengend Snippet: Fig. 4 The lactylation of YY1 in microglia plays an important role in regulating angiogenesis. a Heatmap shows the top 20 DELPs with increased and decreased lactylation under hypoxia for 24 h. b The enrichment analysis of DELPs. c Using the STRING database, DELPs specifically target the binding process. The node size corresponds to the relative fold-change of lactylation. d Collision-induced dissociation (CID) analysis was used to determine the modification sites. The MS/MS spectrum of modified “(Kla)SYLSGGA GAAGGGGADPGNK” is shown. e Hypoxia increases YY1 lactylation. Lactylation of YY1 in HMC3 cells under normoxia or hypoxia for 24 h was detected by YY1-Kla antibody (n = 3 per group). f–i HMC3 cells overexpressing WT or K183R YY1 were pretreated with 1% O2 for 24 h. Then HRMECs were cocultured with the pretreated HMC3 cells. The tube formation, spheroid sprouting, migration, and proliferation assays were performed as shown in the “Methods” (n = 3 independent experiments, 3 images for each group); scale bars, 50 μm (f, g, and i) or 100 μm (h). *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: RNA was isolated using TRIzol reagent from HMC3 cells (1 × 106 cells) or retinal tissue. cDNA was generated by the RT Master Mix for qPCR (MCE, HY-K0510).

Techniques: Binding Assay, Modification, Tandem Mass Spectroscopy, Migration

Fig. 5 YY1 lactylation contributes to angiogenesis by regulating FGF2 expression. a The mRNA expression of VEGFA, FGF2, MMP2, MMP9, and ANGPTL6 in the retinal tissue of normal control (Normal), OIR (DMSO), and OIR (PLX3397) mice (n = 3 per group). b The mRNA expression of VEGFA, FGF2, MMP2, MMP9, and ANGPTL6 in HMC3 cells in the three groups subjected to hypoxia for 0, 12, and 24 h (n = 3 per group). c The protein expression of FGF2 in the retinas of normal control (Normal), OIR (DMSO), and OIR (PLX3397) mice (n = 3 per group). d The protein expression of FGF2 in the HMC3 cells subjected to hypoxia for 0 and 24 h (n = 3 per group). e Representative images of FGF2 co-stained with microglia (Iba1) in retina of OIR mice (OIR P17) and control mice (Normal P17), with quantification of FGF2 intensity (n = the number of Iba1+ cells); scale bar, 20 μm. f The protein expression of FGF2 in HMC3 cells exposed to hypoxia 24 h (Hypoxia Control), hypoxia for 24 h + YY1 WT transfection (Hypoxia + WT), and hypoxia for 24 h + YY1 K183R transfection (Hypoxia + K183R) (n = 3 per group). g ChIP-qPCR analysis of the indicated promoters was performed by using of YY1 antibody in HMC3 cells treated with normoxia and hypoxia (n = 3 per group). h ChIP-qPCR analysis of the indicated promoters was performed by using of YY1 antibody in HMC3 cells overexpressing WT and K183R YY1 (n = 3 per group). i The luciferase activity of the FGF2 promoter driven reporter vector was measured in response to YY1 WT or K183R cotransfection under hypoxia (n = 6 per group). NS, non-significance; *p <0.05; **p < 0.01; ***p < 0.001

Journal: Genome biology

Article Title: YY1 lactylation in microglia promotes angiogenesis through transcription activation-mediated upregulation of FGF2.

doi: 10.1186/s13059-023-02931-y

Figure Lengend Snippet: Fig. 5 YY1 lactylation contributes to angiogenesis by regulating FGF2 expression. a The mRNA expression of VEGFA, FGF2, MMP2, MMP9, and ANGPTL6 in the retinal tissue of normal control (Normal), OIR (DMSO), and OIR (PLX3397) mice (n = 3 per group). b The mRNA expression of VEGFA, FGF2, MMP2, MMP9, and ANGPTL6 in HMC3 cells in the three groups subjected to hypoxia for 0, 12, and 24 h (n = 3 per group). c The protein expression of FGF2 in the retinas of normal control (Normal), OIR (DMSO), and OIR (PLX3397) mice (n = 3 per group). d The protein expression of FGF2 in the HMC3 cells subjected to hypoxia for 0 and 24 h (n = 3 per group). e Representative images of FGF2 co-stained with microglia (Iba1) in retina of OIR mice (OIR P17) and control mice (Normal P17), with quantification of FGF2 intensity (n = the number of Iba1+ cells); scale bar, 20 μm. f The protein expression of FGF2 in HMC3 cells exposed to hypoxia 24 h (Hypoxia Control), hypoxia for 24 h + YY1 WT transfection (Hypoxia + WT), and hypoxia for 24 h + YY1 K183R transfection (Hypoxia + K183R) (n = 3 per group). g ChIP-qPCR analysis of the indicated promoters was performed by using of YY1 antibody in HMC3 cells treated with normoxia and hypoxia (n = 3 per group). h ChIP-qPCR analysis of the indicated promoters was performed by using of YY1 antibody in HMC3 cells overexpressing WT and K183R YY1 (n = 3 per group). i The luciferase activity of the FGF2 promoter driven reporter vector was measured in response to YY1 WT or K183R cotransfection under hypoxia (n = 6 per group). NS, non-significance; *p <0.05; **p < 0.01; ***p < 0.001

Article Snippet: RNA was isolated using TRIzol reagent from HMC3 cells (1 × 106 cells) or retinal tissue. cDNA was generated by the RT Master Mix for qPCR (MCE, HY-K0510).

Techniques: Expressing, Control, Staining, Transfection, ChIP-qPCR, Luciferase, Activity Assay, Plasmid Preparation, Cotransfection

Fig. 6 p300 affects the lactylation of YY1 and then regulates angiogenesis in vitro. a Quantification of p300 in the HMC3 cells under normoxia or hypoxia for 24 h (n = 3 per group). b Co-IP analysis of YY1 and p300. c Colocalization of YY1 and p300 by double-label immunofluorescence. d The Pan-Kla level in the HMC3 cells overexpressing p300 (n = 3 per group). e Overexpression of p300 increased YY1 lactylation. Lactylation of YY1 in control group, or oep300-WT group was detected by YY1-Kla antibodies (n = 3 per group). f The expression of FGF2 and p300 in the HMC3 cells overexpressing p300 (n = 3 per group). g Quantification of FGF2 and Pan-Kla in the HMC3 cells treated with 0, 5, 10, and 20 μM A-485 under hypoxia for 24 h (n = 3 per group). h Applying A-485 decreased YY1 lactylation. Lactylation of YY1 with 0 μM A-485, or 20 μM A-485 was detected by YY1-Kla antibodies (n = 3 per group). *p < 0.05; **p < 0.01; ***p < 0.001

Journal: Genome biology

Article Title: YY1 lactylation in microglia promotes angiogenesis through transcription activation-mediated upregulation of FGF2.

doi: 10.1186/s13059-023-02931-y

Figure Lengend Snippet: Fig. 6 p300 affects the lactylation of YY1 and then regulates angiogenesis in vitro. a Quantification of p300 in the HMC3 cells under normoxia or hypoxia for 24 h (n = 3 per group). b Co-IP analysis of YY1 and p300. c Colocalization of YY1 and p300 by double-label immunofluorescence. d The Pan-Kla level in the HMC3 cells overexpressing p300 (n = 3 per group). e Overexpression of p300 increased YY1 lactylation. Lactylation of YY1 in control group, or oep300-WT group was detected by YY1-Kla antibodies (n = 3 per group). f The expression of FGF2 and p300 in the HMC3 cells overexpressing p300 (n = 3 per group). g Quantification of FGF2 and Pan-Kla in the HMC3 cells treated with 0, 5, 10, and 20 μM A-485 under hypoxia for 24 h (n = 3 per group). h Applying A-485 decreased YY1 lactylation. Lactylation of YY1 with 0 μM A-485, or 20 μM A-485 was detected by YY1-Kla antibodies (n = 3 per group). *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: RNA was isolated using TRIzol reagent from HMC3 cells (1 × 106 cells) or retinal tissue. cDNA was generated by the RT Master Mix for qPCR (MCE, HY-K0510).

Techniques: In Vitro, Co-Immunoprecipitation Assay, Immunofluorescence, Over Expression, Control, Expressing

(A) Schematic outline of the cell culture. Three biological replicates were established for HMC3, HBEC5i and NHA. All cells were grown in DMEM-HG medium under serum-deprived conditions for 24 h after which the media was collected and pooled from the three brain cell types to treat the SKBR3 cells for another 24 h. SKBR3-control cells were maintained in serum-free media for 48 h. (B) Phase contrast microscopy image of non-treated SKBR3 cells under 20X magnification. (C) Immunofluorescence microscopy of non-treated SKBR3 cells labeled with anti-HER2 antibody and detected using an anti-mouse IgG secondary antibody conjugated to CruzFluor488. (D) Phase contrast microscopy image of CM-treated SKBR3 cells under 20X magnification shows development of filopodia/lamellipodia type protrusions from the surface of the cells. (E) Immunofluorescence microscopy of treated SKBR3 cells labeled with anti-HER2 antibody and detected using an anti-mouse IgG secondary antibody conjugated to CruzFluor488. (F) Stacked bar chart of percentage cells in the G1, S and G2 phases of the cell cycle for the control (C) and treated (T) SKBR3 cells.

Journal: bioRxiv

Article Title: Proteomic Insights into Metastatic Breast Cancer Response to Brain Cell-Secreted Factors

doi: 10.1101/2023.10.22.563488

Figure Lengend Snippet: (A) Schematic outline of the cell culture. Three biological replicates were established for HMC3, HBEC5i and NHA. All cells were grown in DMEM-HG medium under serum-deprived conditions for 24 h after which the media was collected and pooled from the three brain cell types to treat the SKBR3 cells for another 24 h. SKBR3-control cells were maintained in serum-free media for 48 h. (B) Phase contrast microscopy image of non-treated SKBR3 cells under 20X magnification. (C) Immunofluorescence microscopy of non-treated SKBR3 cells labeled with anti-HER2 antibody and detected using an anti-mouse IgG secondary antibody conjugated to CruzFluor488. (D) Phase contrast microscopy image of CM-treated SKBR3 cells under 20X magnification shows development of filopodia/lamellipodia type protrusions from the surface of the cells. (E) Immunofluorescence microscopy of treated SKBR3 cells labeled with anti-HER2 antibody and detected using an anti-mouse IgG secondary antibody conjugated to CruzFluor488. (F) Stacked bar chart of percentage cells in the G1, S and G2 phases of the cell cycle for the control (C) and treated (T) SKBR3 cells.

Article Snippet: Human HER2+ breast cancer cells (SKBR3), microglia (HMC3), brain endothelial cells (HBEC5i), and penicillin-streptomycin (PenStrep) solution were purchased from ATCC (Manassas, VA, USA).

Techniques: Cell Culture, Control, Microscopy, Immunofluorescence, Labeling

A549 ( A ) and HCM3 ( B ) cells were transfected for 24 h with pcDNA3 expressing rNS4B protein from ZIKV-15555 (rNS4B ZIKV-15555 ), MR766 (rNS4B MR766 ), or BeH819015 (rNS4B BeH819015 ). The rNS4B ZIK-15555 protein mutant lacking the 2K peptide is indicated as (Δ2K)-rNS4B ZIK-15555 . Cell lysates in RIPA buffer were assessed by immunoblot assays with anti-epitope antibody FLAG as the primary antibody. The β-tubulin protein served as a protein-loading control. The positions of rNS4B are indicated.

Journal: PLOS Neglected Tropical Diseases

Article Title: Replication properties of a contemporary Zika virus from West Africa

doi: 10.1371/journal.pntd.0012066

Figure Lengend Snippet: A549 ( A ) and HCM3 ( B ) cells were transfected for 24 h with pcDNA3 expressing rNS4B protein from ZIKV-15555 (rNS4B ZIKV-15555 ), MR766 (rNS4B MR766 ), or BeH819015 (rNS4B BeH819015 ). The rNS4B ZIK-15555 protein mutant lacking the 2K peptide is indicated as (Δ2K)-rNS4B ZIK-15555 . Cell lysates in RIPA buffer were assessed by immunoblot assays with anti-epitope antibody FLAG as the primary antibody. The β-tubulin protein served as a protein-loading control. The positions of rNS4B are indicated.

Article Snippet: Human embryonic kidney HEK-293T (ATCC, CRL-1573), human carcinoma epithelial lung A549 (Invivogen Inc, Toulouse, France), human microglial clone 3 HCM3 (ATCC, CCL-3304) and monkey kidney VeroE6 (CCL-81, ATCC, Manassas, VA, USA) cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) growth medium supplemented with 10% (HEK-293T, A549 and HCM3 cells) or 5% (VeroE6 cells) of heat-inactivated fetal bovine serum (FBS, Dutscher, Brumath, France), and antibiotics (PAN Biotech Dutscher, Brumath, France) at 37°C under a 5% CO 2 atmosphere.

Techniques: Transfection, Expressing, Mutagenesis, Western Blot, Control