hbmec cells Search Results


gp96  (StressMarq)
93
StressMarq gp96
HSP90B1 and HSPA5 genes and <t>GP96</t> are induced in ALD. (A) mRNA abundance of HSP90B1 and HSPA5 genes (in transcripts per million) was evaluated by RNA sequencing in patients with normal liver (n = 10), early ASH (n = 12), severe_AH (n = 17), explants_AH (n = 10), and livers from NAFLD (n = 10), HCV (n = 10), and comp_Cirrhosis (n = 9). (B) Representative immunohistochemistry of GP96 in human normal and alcoholic liver (magnification × 200). WT mice were fed with control liquid diet (pair‐fed) or diet containing 5% alcohol for 4 weeks. The mRNA expression of GP96 and GRP78 was measured by RT‐PCR in livers (C) and isolated hepatocytes (E) and liver macrophages (n = 4‐10). (D) Protein level of GP96 and GRP78 in liver lysate was analyzed by western blotting (n = 6). (F) BMDMs were treated with 25 mM ethanol and mRNA expression of GP96 and GRP78 (n = 6). (G) The protein level of GP96 was analyzed at different time intervals (n = 3). Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Abbreviations: EtOH, alcohol; comp, compensated; tpm, transcripts per million.
Gp96, supplied by StressMarq, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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grp94  (Boster Bio)
91
Boster Bio grp94
HSP90B1 and HSPA5 genes and <t>GP96</t> are induced in ALD. (A) mRNA abundance of HSP90B1 and HSPA5 genes (in transcripts per million) was evaluated by RNA sequencing in patients with normal liver (n = 10), early ASH (n = 12), severe_AH (n = 17), explants_AH (n = 10), and livers from NAFLD (n = 10), HCV (n = 10), and comp_Cirrhosis (n = 9). (B) Representative immunohistochemistry of GP96 in human normal and alcoholic liver (magnification × 200). WT mice were fed with control liquid diet (pair‐fed) or diet containing 5% alcohol for 4 weeks. The mRNA expression of GP96 and GRP78 was measured by RT‐PCR in livers (C) and isolated hepatocytes (E) and liver macrophages (n = 4‐10). (D) Protein level of GP96 and GRP78 in liver lysate was analyzed by western blotting (n = 6). (F) BMDMs were treated with 25 mM ethanol and mRNA expression of GP96 and GRP78 (n = 6). (G) The protein level of GP96 was analyzed at different time intervals (n = 3). Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Abbreviations: EtOH, alcohol; comp, compensated; tpm, transcripts per million.
Grp94, supplied by Boster Bio, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 91 stars, based on 1 article reviews
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grp75  (StressMarq)
91
StressMarq grp75
HSP90B1 and HSPA5 genes and <t>GP96</t> are induced in ALD. (A) mRNA abundance of HSP90B1 and HSPA5 genes (in transcripts per million) was evaluated by RNA sequencing in patients with normal liver (n = 10), early ASH (n = 12), severe_AH (n = 17), explants_AH (n = 10), and livers from NAFLD (n = 10), HCV (n = 10), and comp_Cirrhosis (n = 9). (B) Representative immunohistochemistry of GP96 in human normal and alcoholic liver (magnification × 200). WT mice were fed with control liquid diet (pair‐fed) or diet containing 5% alcohol for 4 weeks. The mRNA expression of GP96 and GRP78 was measured by RT‐PCR in livers (C) and isolated hepatocytes (E) and liver macrophages (n = 4‐10). (D) Protein level of GP96 and GRP78 in liver lysate was analyzed by western blotting (n = 6). (F) BMDMs were treated with 25 mM ethanol and mRNA expression of GP96 and GRP78 (n = 6). (G) The protein level of GP96 was analyzed at different time intervals (n = 3). Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Abbreviations: EtOH, alcohol; comp, compensated; tpm, transcripts per million.
Grp75, supplied by StressMarq, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human brain microvascular endothelial cells  (ScienCell)
90
ScienCell human brain microvascular endothelial cells
( A ) CHIKV-infected <t>HBMEC</t> cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.
Human Brain Microvascular Endothelial Cells, supplied by ScienCell, 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 brain microvascular endothelial cells - by Bioz Stars, 2026-03
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hmec-1  (Sanquin)
90
Sanquin hmec-1
( A ) CHIKV-infected <t>HBMEC</t> cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.
Hmec 1, supplied by Sanquin, 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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hmec-1 - by Bioz Stars, 2026-03
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hbmecs  (Cell Systems Corporation)
90
Cell Systems Corporation hbmecs
( A ) CHIKV-infected <t>HBMEC</t> cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.
Hbmecs, supplied by Cell Systems Corporation, 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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Average 90 stars, based on 1 article reviews
hbmecs - by Bioz Stars, 2026-03
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primary hbmecs acbri376  (Cell Systems Corporation)
90
Cell Systems Corporation primary hbmecs acbri376
( A ) CHIKV-infected <t>HBMEC</t> cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.
Primary Hbmecs Acbri376, supplied by Cell Systems Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/primary hbmecs acbri376/product/Cell Systems Corporation
Average 90 stars, based on 1 article reviews
primary hbmecs acbri376 - by Bioz Stars, 2026-03
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immortalized human brain microvascular endothelial cells, hbmec  (Johns Hopkins HealthCare)
90
Johns Hopkins HealthCare immortalized human brain microvascular endothelial cells, hbmec
( A ) CHIKV-infected <t>HBMEC</t> cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.
Immortalized Human Brain Microvascular Endothelial Cells, Hbmec, supplied by Johns Hopkins HealthCare, 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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Average 90 stars, based on 1 article reviews
immortalized human brain microvascular endothelial cells, hbmec - by Bioz Stars, 2026-03
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human brain microvascular endothelial cells (hbmecs) icell- h070  (iCell Gene Therapeutics)
90
iCell Gene Therapeutics human brain microvascular endothelial cells (hbmecs) icell- h070
( A ) CHIKV-infected <t>HBMEC</t> cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.
Human Brain Microvascular Endothelial Cells (Hbmecs) Icell H070, supplied by iCell Gene Therapeutics, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human brain microvascular endothelial cells (hbmecs) icell- h070/product/iCell Gene Therapeutics
Average 90 stars, based on 1 article reviews
human brain microvascular endothelial cells (hbmecs) icell- h070 - by Bioz Stars, 2026-03
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human brain-microvascular endothelial cells (hbmecs)  (Biocompare)
90
Biocompare human brain-microvascular endothelial cells (hbmecs)
( A ) CHIKV-infected <t>HBMEC</t> cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.
Human Brain Microvascular Endothelial Cells (Hbmecs), supplied by Biocompare, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human brain-microvascular endothelial cells (hbmecs)/product/Biocompare
Average 90 stars, based on 1 article reviews
human brain-microvascular endothelial cells (hbmecs) - by Bioz Stars, 2026-03
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transfected human bmec  (Johns Hopkins HealthCare)
90
Johns Hopkins HealthCare transfected human bmec
( A ) CHIKV-infected <t>HBMEC</t> cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.
Transfected Human Bmec, supplied by Johns Hopkins HealthCare, 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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Average 90 stars, based on 1 article reviews
transfected human bmec - by Bioz Stars, 2026-03
90/100 stars
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hbmecs acbri 376  (Cell Systems Corporation)
90
Cell Systems Corporation hbmecs acbri 376
( A ) CHIKV-infected <t>HBMEC</t> cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.
Hbmecs Acbri 376, supplied by Cell Systems Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/hbmecs acbri 376/product/Cell Systems Corporation
Average 90 stars, based on 1 article reviews
hbmecs acbri 376 - by Bioz Stars, 2026-03
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Image Search Results


HSP90B1 and HSPA5 genes and GP96 are induced in ALD. (A) mRNA abundance of HSP90B1 and HSPA5 genes (in transcripts per million) was evaluated by RNA sequencing in patients with normal liver (n = 10), early ASH (n = 12), severe_AH (n = 17), explants_AH (n = 10), and livers from NAFLD (n = 10), HCV (n = 10), and comp_Cirrhosis (n = 9). (B) Representative immunohistochemistry of GP96 in human normal and alcoholic liver (magnification × 200). WT mice were fed with control liquid diet (pair‐fed) or diet containing 5% alcohol for 4 weeks. The mRNA expression of GP96 and GRP78 was measured by RT‐PCR in livers (C) and isolated hepatocytes (E) and liver macrophages (n = 4‐10). (D) Protein level of GP96 and GRP78 in liver lysate was analyzed by western blotting (n = 6). (F) BMDMs were treated with 25 mM ethanol and mRNA expression of GP96 and GRP78 (n = 6). (G) The protein level of GP96 was analyzed at different time intervals (n = 3). Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Abbreviations: EtOH, alcohol; comp, compensated; tpm, transcripts per million.

Journal: Hepatology Communications

Article Title: Myeloid Endoplasmic Reticulum Resident Chaperone GP96 Facilitates Inflammation and Steatosis in Alcohol‐Associated Liver Disease

doi: 10.1002/hep4.1713

Figure Lengend Snippet: HSP90B1 and HSPA5 genes and GP96 are induced in ALD. (A) mRNA abundance of HSP90B1 and HSPA5 genes (in transcripts per million) was evaluated by RNA sequencing in patients with normal liver (n = 10), early ASH (n = 12), severe_AH (n = 17), explants_AH (n = 10), and livers from NAFLD (n = 10), HCV (n = 10), and comp_Cirrhosis (n = 9). (B) Representative immunohistochemistry of GP96 in human normal and alcoholic liver (magnification × 200). WT mice were fed with control liquid diet (pair‐fed) or diet containing 5% alcohol for 4 weeks. The mRNA expression of GP96 and GRP78 was measured by RT‐PCR in livers (C) and isolated hepatocytes (E) and liver macrophages (n = 4‐10). (D) Protein level of GP96 and GRP78 in liver lysate was analyzed by western blotting (n = 6). (F) BMDMs were treated with 25 mM ethanol and mRNA expression of GP96 and GRP78 (n = 6). (G) The protein level of GP96 was analyzed at different time intervals (n = 3). Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Abbreviations: EtOH, alcohol; comp, compensated; tpm, transcripts per million.

Article Snippet: Antibodies used were GP96 (SMC‐105; StressMarq Biosciences, Cadboro, Canada), GRP78 (CST‐3183; Cell Signaling Technology), peroxisome proliferator‐activated receptor alpha (PPAR‐α) (sc‐9000; Santa Cruz Biotechnology, Dallas, TX), CyP2e1 (AB1252; MilliporeSigma, Burlington, MA), calnexin (ab2295; Abcam, Cambridge, United Kingdom), C/EBP‐homologous protein (CHOP) (sc‐7351; Santa Cruz Biotechnology), ATF6α (sc‐166659; Santa Cruz Biotechnology), ATF4 (10835‐1‐AP; Proteintech, Rosemont, IL), ATF3 (sc‐81189; Santa Cruz Biotechnology), β‐actin (ab6276; Abcam), tubulin (ab6046; Abcam), and TATA‐box protein (TBP) (ab818, Abcam).

Techniques: RNA Sequencing Assay, Immunohistochemistry, Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Western Blot

Myeloid‐specific GP96 deficiency alleviates chronic alcohol‐induced liver injury. Female WT and M‐GP96KO mice were fed with isocaloric control liquid diet (pair‐fed) or 5% alcohol‐containing Leiber‐DeCarli diet (alcohol‐fed) for 4 weeks, and liver‐to–body weight ratio (A), serum levels of ALT (n = 10‐16) (B), and liver triglycerides (C) were analyzed. Representative photomicrographs of hepatic injury and steatosis assessed by H&E (D) and Oil Red O (E) staining in livers of mice of the indicated genotype (magnification × 100). Percentage of Oil Red O–positive area was quantitated by ImageJ (n = 3‐6). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Abbreviations: EtOH, alcohol; wt, weight.

Journal: Hepatology Communications

Article Title: Myeloid Endoplasmic Reticulum Resident Chaperone GP96 Facilitates Inflammation and Steatosis in Alcohol‐Associated Liver Disease

doi: 10.1002/hep4.1713

Figure Lengend Snippet: Myeloid‐specific GP96 deficiency alleviates chronic alcohol‐induced liver injury. Female WT and M‐GP96KO mice were fed with isocaloric control liquid diet (pair‐fed) or 5% alcohol‐containing Leiber‐DeCarli diet (alcohol‐fed) for 4 weeks, and liver‐to–body weight ratio (A), serum levels of ALT (n = 10‐16) (B), and liver triglycerides (C) were analyzed. Representative photomicrographs of hepatic injury and steatosis assessed by H&E (D) and Oil Red O (E) staining in livers of mice of the indicated genotype (magnification × 100). Percentage of Oil Red O–positive area was quantitated by ImageJ (n = 3‐6). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Abbreviations: EtOH, alcohol; wt, weight.

Article Snippet: Antibodies used were GP96 (SMC‐105; StressMarq Biosciences, Cadboro, Canada), GRP78 (CST‐3183; Cell Signaling Technology), peroxisome proliferator‐activated receptor alpha (PPAR‐α) (sc‐9000; Santa Cruz Biotechnology, Dallas, TX), CyP2e1 (AB1252; MilliporeSigma, Burlington, MA), calnexin (ab2295; Abcam, Cambridge, United Kingdom), C/EBP‐homologous protein (CHOP) (sc‐7351; Santa Cruz Biotechnology), ATF6α (sc‐166659; Santa Cruz Biotechnology), ATF4 (10835‐1‐AP; Proteintech, Rosemont, IL), ATF3 (sc‐81189; Santa Cruz Biotechnology), β‐actin (ab6276; Abcam), tubulin (ab6046; Abcam), and TATA‐box protein (TBP) (ab818, Abcam).

Techniques: Staining

Mice lacking myeloid‐specific GP96 exhibit altered lipid metabolism after 4 weeks of alcohol consumption. (A) PPAR‐α protein was detected in nuclear extracts of livers by western blot, and expression was quantitated and normalized to pair‐fed group (n = 4). TBP was used as loading control. The mRNA expression of genes involved in fatty acid β‐oxidation (CPT1a, ACOX1, LCAD, and MCAD) (B‐E) and lipogenesis (SREBPF1, SCD1, and FAS) (F‐H) was analyzed in WT and M‐GP96KO hepatocytes by RT‐PCR and compared with pair‐fed control (n = 3‐6). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Journal: Hepatology Communications

Article Title: Myeloid Endoplasmic Reticulum Resident Chaperone GP96 Facilitates Inflammation and Steatosis in Alcohol‐Associated Liver Disease

doi: 10.1002/hep4.1713

Figure Lengend Snippet: Mice lacking myeloid‐specific GP96 exhibit altered lipid metabolism after 4 weeks of alcohol consumption. (A) PPAR‐α protein was detected in nuclear extracts of livers by western blot, and expression was quantitated and normalized to pair‐fed group (n = 4). TBP was used as loading control. The mRNA expression of genes involved in fatty acid β‐oxidation (CPT1a, ACOX1, LCAD, and MCAD) (B‐E) and lipogenesis (SREBPF1, SCD1, and FAS) (F‐H) was analyzed in WT and M‐GP96KO hepatocytes by RT‐PCR and compared with pair‐fed control (n = 3‐6). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Antibodies used were GP96 (SMC‐105; StressMarq Biosciences, Cadboro, Canada), GRP78 (CST‐3183; Cell Signaling Technology), peroxisome proliferator‐activated receptor alpha (PPAR‐α) (sc‐9000; Santa Cruz Biotechnology, Dallas, TX), CyP2e1 (AB1252; MilliporeSigma, Burlington, MA), calnexin (ab2295; Abcam, Cambridge, United Kingdom), C/EBP‐homologous protein (CHOP) (sc‐7351; Santa Cruz Biotechnology), ATF6α (sc‐166659; Santa Cruz Biotechnology), ATF4 (10835‐1‐AP; Proteintech, Rosemont, IL), ATF3 (sc‐81189; Santa Cruz Biotechnology), β‐actin (ab6276; Abcam), tubulin (ab6046; Abcam), and TATA‐box protein (TBP) (ab818, Abcam).

Techniques: Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction

Myeloid‐specific GP96 deficiency prevents chronic alcohol‐induced endotoxin and pro‐inflammatory cytokine production. (A) Endotoxin level was measured in serum (n = 8‐12). (B) CyP2e1 was detected in liver microsomal fraction by western blot, and calnexin was used as an internal loading control (n = 3). Total RNA from liver tissue was subjected to quantitative RT‐PCR for analysis of pro‐inflammatory cytokines TNF‐α, IL‐6, MCP‐1, and IL‐1β; NLRP3; anti‐inflammatory markers IL‐10, TGF‐β, and ATF3 (C); and macrophage markers (n = 6‐10) (F). (D) Total liver protein level for TNF‐α was measured in tissue extracts of pair‐fed and alcohol‐fed mice by ELISA (n = 6‐10). (E) ATF3 protein level was analyzed by western blot in whole‐liver extracts using tubulin as an internal control (n = 3). The mRNA expression profile of (G) pro‐inflammatory, and (H) anti‐inflammatory and restorative macrophage markers, Trem2 and ATF3, were analyzed in liver macrophages of pair‐fed and alcohol‐fed mice (n = 5). (I) The mRNA level of pro‐inflammatory cytokines was analyzed in BMDMs isolated from WT and M‐GP96KO mice and stimulated with 100 ng/mL LPS for 2 hours (n = 9). Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Journal: Hepatology Communications

Article Title: Myeloid Endoplasmic Reticulum Resident Chaperone GP96 Facilitates Inflammation and Steatosis in Alcohol‐Associated Liver Disease

doi: 10.1002/hep4.1713

Figure Lengend Snippet: Myeloid‐specific GP96 deficiency prevents chronic alcohol‐induced endotoxin and pro‐inflammatory cytokine production. (A) Endotoxin level was measured in serum (n = 8‐12). (B) CyP2e1 was detected in liver microsomal fraction by western blot, and calnexin was used as an internal loading control (n = 3). Total RNA from liver tissue was subjected to quantitative RT‐PCR for analysis of pro‐inflammatory cytokines TNF‐α, IL‐6, MCP‐1, and IL‐1β; NLRP3; anti‐inflammatory markers IL‐10, TGF‐β, and ATF3 (C); and macrophage markers (n = 6‐10) (F). (D) Total liver protein level for TNF‐α was measured in tissue extracts of pair‐fed and alcohol‐fed mice by ELISA (n = 6‐10). (E) ATF3 protein level was analyzed by western blot in whole‐liver extracts using tubulin as an internal control (n = 3). The mRNA expression profile of (G) pro‐inflammatory, and (H) anti‐inflammatory and restorative macrophage markers, Trem2 and ATF3, were analyzed in liver macrophages of pair‐fed and alcohol‐fed mice (n = 5). (I) The mRNA level of pro‐inflammatory cytokines was analyzed in BMDMs isolated from WT and M‐GP96KO mice and stimulated with 100 ng/mL LPS for 2 hours (n = 9). Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Antibodies used were GP96 (SMC‐105; StressMarq Biosciences, Cadboro, Canada), GRP78 (CST‐3183; Cell Signaling Technology), peroxisome proliferator‐activated receptor alpha (PPAR‐α) (sc‐9000; Santa Cruz Biotechnology, Dallas, TX), CyP2e1 (AB1252; MilliporeSigma, Burlington, MA), calnexin (ab2295; Abcam, Cambridge, United Kingdom), C/EBP‐homologous protein (CHOP) (sc‐7351; Santa Cruz Biotechnology), ATF6α (sc‐166659; Santa Cruz Biotechnology), ATF4 (10835‐1‐AP; Proteintech, Rosemont, IL), ATF3 (sc‐81189; Santa Cruz Biotechnology), β‐actin (ab6276; Abcam), tubulin (ab6046; Abcam), and TATA‐box protein (TBP) (ab818, Abcam).

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

Loss of myeloid‐specific GP96 prevents LPS‐induced liver injury and inflammation. Female WT and M‐GP96KO mice were injected intraperitoneally with LPS (0.5 mg/kg body weight or saline (n = 4‐8). Serum was collected after 18 hours and subjected to analysis of ALT (A) and AST (B) and compared with the control group. (C) Livers were collected 2 hours after LPS injection, and liver cytokine mRNA was analyzed by RT‐PCR. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Journal: Hepatology Communications

Article Title: Myeloid Endoplasmic Reticulum Resident Chaperone GP96 Facilitates Inflammation and Steatosis in Alcohol‐Associated Liver Disease

doi: 10.1002/hep4.1713

Figure Lengend Snippet: Loss of myeloid‐specific GP96 prevents LPS‐induced liver injury and inflammation. Female WT and M‐GP96KO mice were injected intraperitoneally with LPS (0.5 mg/kg body weight or saline (n = 4‐8). Serum was collected after 18 hours and subjected to analysis of ALT (A) and AST (B) and compared with the control group. (C) Livers were collected 2 hours after LPS injection, and liver cytokine mRNA was analyzed by RT‐PCR. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Article Snippet: Antibodies used were GP96 (SMC‐105; StressMarq Biosciences, Cadboro, Canada), GRP78 (CST‐3183; Cell Signaling Technology), peroxisome proliferator‐activated receptor alpha (PPAR‐α) (sc‐9000; Santa Cruz Biotechnology, Dallas, TX), CyP2e1 (AB1252; MilliporeSigma, Burlington, MA), calnexin (ab2295; Abcam, Cambridge, United Kingdom), C/EBP‐homologous protein (CHOP) (sc‐7351; Santa Cruz Biotechnology), ATF6α (sc‐166659; Santa Cruz Biotechnology), ATF4 (10835‐1‐AP; Proteintech, Rosemont, IL), ATF3 (sc‐81189; Santa Cruz Biotechnology), β‐actin (ab6276; Abcam), tubulin (ab6046; Abcam), and TATA‐box protein (TBP) (ab818, Abcam).

Techniques: Injection, Reverse Transcription Polymerase Chain Reaction

Inhibition of GP96 using specific inhibitor PU‐WS13 and siRNA reduces LPS‐induced pro‐inflammatory cytokine production. BMDMs were isolated from C57BL/6J mice and stimulated with LPS (100 ng/mL) for 2 hours and treated with PU‐WS13 (0.5 μM) either alone for 2 hours or before LPS for 1 hour. DMSO‐treated cells served as control group (n = 8). RT‐PCR was carried out to evaluate the expression of TNF‐α (A), IL‐6 (B), IL‐1β (C), and MCP‐1 (D) and compared with the untreated group. (E) Culture supernatant was evaluated for secreted TNF‐α by ELISA. (F) Murine macrophage RAW 264.7 cell line was transiently transfected with GP96 siRNA (100 nM) or negative control siRNA for 48 hours and stimulated with LPS for the final 6 hours. Secreted TNF‐α level was measured in culture supernatant by ELISA. Data are presented as mean ± SEM. *** P < 0.001, **** P < 0.0001. Abbreviation: ND, not detected.

Journal: Hepatology Communications

Article Title: Myeloid Endoplasmic Reticulum Resident Chaperone GP96 Facilitates Inflammation and Steatosis in Alcohol‐Associated Liver Disease

doi: 10.1002/hep4.1713

Figure Lengend Snippet: Inhibition of GP96 using specific inhibitor PU‐WS13 and siRNA reduces LPS‐induced pro‐inflammatory cytokine production. BMDMs were isolated from C57BL/6J mice and stimulated with LPS (100 ng/mL) for 2 hours and treated with PU‐WS13 (0.5 μM) either alone for 2 hours or before LPS for 1 hour. DMSO‐treated cells served as control group (n = 8). RT‐PCR was carried out to evaluate the expression of TNF‐α (A), IL‐6 (B), IL‐1β (C), and MCP‐1 (D) and compared with the untreated group. (E) Culture supernatant was evaluated for secreted TNF‐α by ELISA. (F) Murine macrophage RAW 264.7 cell line was transiently transfected with GP96 siRNA (100 nM) or negative control siRNA for 48 hours and stimulated with LPS for the final 6 hours. Secreted TNF‐α level was measured in culture supernatant by ELISA. Data are presented as mean ± SEM. *** P < 0.001, **** P < 0.0001. Abbreviation: ND, not detected.

Article Snippet: Antibodies used were GP96 (SMC‐105; StressMarq Biosciences, Cadboro, Canada), GRP78 (CST‐3183; Cell Signaling Technology), peroxisome proliferator‐activated receptor alpha (PPAR‐α) (sc‐9000; Santa Cruz Biotechnology, Dallas, TX), CyP2e1 (AB1252; MilliporeSigma, Burlington, MA), calnexin (ab2295; Abcam, Cambridge, United Kingdom), C/EBP‐homologous protein (CHOP) (sc‐7351; Santa Cruz Biotechnology), ATF6α (sc‐166659; Santa Cruz Biotechnology), ATF4 (10835‐1‐AP; Proteintech, Rosemont, IL), ATF3 (sc‐81189; Santa Cruz Biotechnology), β‐actin (ab6276; Abcam), tubulin (ab6046; Abcam), and TATA‐box protein (TBP) (ab818, Abcam).

Techniques: Inhibition, Isolation, Reverse Transcription Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Transfection, Negative Control

Schematic representation depicting pathophysiological significance of macrophage‐specific GP96 during chronic alcohol‐mediated liver inflammation and injury.

Journal: Hepatology Communications

Article Title: Myeloid Endoplasmic Reticulum Resident Chaperone GP96 Facilitates Inflammation and Steatosis in Alcohol‐Associated Liver Disease

doi: 10.1002/hep4.1713

Figure Lengend Snippet: Schematic representation depicting pathophysiological significance of macrophage‐specific GP96 during chronic alcohol‐mediated liver inflammation and injury.

Article Snippet: Antibodies used were GP96 (SMC‐105; StressMarq Biosciences, Cadboro, Canada), GRP78 (CST‐3183; Cell Signaling Technology), peroxisome proliferator‐activated receptor alpha (PPAR‐α) (sc‐9000; Santa Cruz Biotechnology, Dallas, TX), CyP2e1 (AB1252; MilliporeSigma, Burlington, MA), calnexin (ab2295; Abcam, Cambridge, United Kingdom), C/EBP‐homologous protein (CHOP) (sc‐7351; Santa Cruz Biotechnology), ATF6α (sc‐166659; Santa Cruz Biotechnology), ATF4 (10835‐1‐AP; Proteintech, Rosemont, IL), ATF3 (sc‐81189; Santa Cruz Biotechnology), β‐actin (ab6276; Abcam), tubulin (ab6046; Abcam), and TATA‐box protein (TBP) (ab818, Abcam).

Techniques:

( A ) CHIKV-infected HBMEC cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.

Journal: PLoS Neglected Tropical Diseases

Article Title: A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

doi: 10.1371/journal.pntd.0002661

Figure Lengend Snippet: ( A ) CHIKV-infected HBMEC cells were viewed under DIC microscopy to observe for morphological changes. The cell density was lower in CHIKV-infected cells as compared to mock-infected cells from 48 h.p.i. onwards. ( B ) CHIKV-infected HBMEC cells were fixed at various intervals post-infection and immunofluorescence assay was performed to detect for CHIKV protein expression (green). Cell nuclei were stained with DAPI (blue). Moderate amounts of CHIKV protein expression were observed at 24 h.p.i. of HBMEC. ( C ) Quantification of infectious virus titer by plaque assay showed an increasing trend, with a peak in infectious virus titer at 36 h.p.i.. Vertical bars represent one standard deviation from the mean of three readings.

Article Snippet: Human Brain Microvascular Endothelial Cells (HBMEC) from ScienCell were maintained in Endothelial Cell Medium (ECM) supplemented with 5% fetal bovine serum (FBS) and 1% endothelial cell growth supplement.

Techniques: Infection, Microscopy, Immunofluorescence, Expressing, Staining, Virus, Plaque Assay, Standard Deviation

( A ) TEER measurements of Vero, Vero C1008 and HBMEC cell monolayers were taken at 24 h.p.i.. The TEER measurements post-apical and basolateral infection were comparable to that of mock-infected cells. Vertical bars represent one standard deviation from the mean of three readings. ( B ) Immunofluorescence assays demonstrated the expression of ZO-1 tight junction proteins (green) in apically-infected and basolaterally-infected Vero, Vero C1008 and HBMEC cells at 24 h.p.i.. The expression of ZO-1 proteins in infected cells is comparable to that of mock-infected cells. (C) The FITC-dextran permeability assays demonstrated that the integrity of Vero and Vero C1008 cell monolayers remained intact at 24 h.p.i., where the permeability of the cell monolayers to FITC-dextran remained low as compared to the TNF-treated cells.

Journal: PLoS Neglected Tropical Diseases

Article Title: A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

doi: 10.1371/journal.pntd.0002661

Figure Lengend Snippet: ( A ) TEER measurements of Vero, Vero C1008 and HBMEC cell monolayers were taken at 24 h.p.i.. The TEER measurements post-apical and basolateral infection were comparable to that of mock-infected cells. Vertical bars represent one standard deviation from the mean of three readings. ( B ) Immunofluorescence assays demonstrated the expression of ZO-1 tight junction proteins (green) in apically-infected and basolaterally-infected Vero, Vero C1008 and HBMEC cells at 24 h.p.i.. The expression of ZO-1 proteins in infected cells is comparable to that of mock-infected cells. (C) The FITC-dextran permeability assays demonstrated that the integrity of Vero and Vero C1008 cell monolayers remained intact at 24 h.p.i., where the permeability of the cell monolayers to FITC-dextran remained low as compared to the TNF-treated cells.

Article Snippet: Human Brain Microvascular Endothelial Cells (HBMEC) from ScienCell were maintained in Endothelial Cell Medium (ECM) supplemented with 5% fetal bovine serum (FBS) and 1% endothelial cell growth supplement.

Techniques: Infection, Standard Deviation, Immunofluorescence, Expressing, Permeability

( A ) The total virus yield at 24 h.p.i. of non-polarized Vero, polarized Vero C1008 and polarized HBMEC cells at an MOI of 10 was quantified by viral plaque assay. Entry of CHIKV is bi-directional in non-polarized Vero cells but occurs preferentially at the apical domain of polarized Vero C1008 and HBMEC cells. Two-tailed Student's t -test: * p<0.05, ** p<0.005. Vertical bars represent one standard deviation from the mean of three readings. ( B ) Virus binding assays show higher amount of CHIKV binding upon apical infection as compared to upon basolateral infection of polarized Vero C1008 cells. In contrast, the amount of CHIKV binding upon apical and basolateral infection of non-polarized Vero cells is similar. Two-tailed Student's t -test: *** p<0.001.

Journal: PLoS Neglected Tropical Diseases

Article Title: A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

doi: 10.1371/journal.pntd.0002661

Figure Lengend Snippet: ( A ) The total virus yield at 24 h.p.i. of non-polarized Vero, polarized Vero C1008 and polarized HBMEC cells at an MOI of 10 was quantified by viral plaque assay. Entry of CHIKV is bi-directional in non-polarized Vero cells but occurs preferentially at the apical domain of polarized Vero C1008 and HBMEC cells. Two-tailed Student's t -test: * p<0.05, ** p<0.005. Vertical bars represent one standard deviation from the mean of three readings. ( B ) Virus binding assays show higher amount of CHIKV binding upon apical infection as compared to upon basolateral infection of polarized Vero C1008 cells. In contrast, the amount of CHIKV binding upon apical and basolateral infection of non-polarized Vero cells is similar. Two-tailed Student's t -test: *** p<0.001.

Article Snippet: Human Brain Microvascular Endothelial Cells (HBMEC) from ScienCell were maintained in Endothelial Cell Medium (ECM) supplemented with 5% fetal bovine serum (FBS) and 1% endothelial cell growth supplement.

Techniques: Virus, Viral Plaque Assay, Two Tailed Test, Standard Deviation, Binding Assay, Infection

( A ) Infectious virus titer of supernatants collected from the apical and basolateral chambers at 24 h.p.i. of non-polarized Vero, polarized Vero C1008 and polarized HBMEC cells at an MOI of 10 were quantified by viral plaque assays. Release of CHIKV is bi-directional in non-polarized Vero cells but occurs preferentially at the apical domain of polarized Vero C1008 and HBMEC cells. Two-tailed Student's t -test: * p<0.05, ** p<0.005, *** p<0.001. Vertical bars represent one standard deviation from the mean of three readings. ( B ) Apically-infected Vero C1008, ( C ) basolaterally-infected Vero C1008, ( D ) apically-infected Vero and ( E ) basolaterally-infected Vero cells were co-labeled with antibodies against CHIKV E2 glycoprotein (green, arrows) and ZO-1 (red, arrowheads). Cell nuclei were stained with DAPI (blue). Z-stacked images show the polarized release of CHIKV towards the apical plasma membrane of Vero C1008 upon apical and basolateral infection. Release of CHIKV is bidirectional in non-polarized Vero cells.

Journal: PLoS Neglected Tropical Diseases

Article Title: A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

doi: 10.1371/journal.pntd.0002661

Figure Lengend Snippet: ( A ) Infectious virus titer of supernatants collected from the apical and basolateral chambers at 24 h.p.i. of non-polarized Vero, polarized Vero C1008 and polarized HBMEC cells at an MOI of 10 were quantified by viral plaque assays. Release of CHIKV is bi-directional in non-polarized Vero cells but occurs preferentially at the apical domain of polarized Vero C1008 and HBMEC cells. Two-tailed Student's t -test: * p<0.05, ** p<0.005, *** p<0.001. Vertical bars represent one standard deviation from the mean of three readings. ( B ) Apically-infected Vero C1008, ( C ) basolaterally-infected Vero C1008, ( D ) apically-infected Vero and ( E ) basolaterally-infected Vero cells were co-labeled with antibodies against CHIKV E2 glycoprotein (green, arrows) and ZO-1 (red, arrowheads). Cell nuclei were stained with DAPI (blue). Z-stacked images show the polarized release of CHIKV towards the apical plasma membrane of Vero C1008 upon apical and basolateral infection. Release of CHIKV is bidirectional in non-polarized Vero cells.

Article Snippet: Human Brain Microvascular Endothelial Cells (HBMEC) from ScienCell were maintained in Endothelial Cell Medium (ECM) supplemented with 5% fetal bovine serum (FBS) and 1% endothelial cell growth supplement.

Techniques: Virus, Two Tailed Test, Standard Deviation, Infection, Labeling, Staining, Clinical Proteomics, Membrane