105 cells Search Results


cd11b c fitc  (Miltenyi Biotec)
98
Miltenyi Biotec cd11b c fitc
Cd11b C Fitc, supplied by Miltenyi Biotec, 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/105+cells/pmc06896275-34-35-41?v=Miltenyi+Biotec
Average 98 stars, based on 1 article reviews
cd11b c fitc - by Bioz Stars, 2026-08
98/100 stars
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detection kit  (Miltenyi Biotec)
90
Miltenyi Biotec detection kit
Detection Kit, supplied by Miltenyi Biotec, 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/product/105+cells/bio_rxiv__2020__01__21__914663-57-19-22?v=Miltenyi+Biotec
Average 90 stars, based on 1 article reviews
detection kit - by Bioz Stars, 2026-08
90/100 stars
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anti cd23 microbeads  (Miltenyi Biotec)
97
Miltenyi Biotec anti cd23 microbeads
Anti Cd23 Microbeads, supplied by Miltenyi Biotec, 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/105+cells/pm37095377-558-15-18?v=Miltenyi+Biotec
Average 97 stars, based on 1 article reviews
anti cd23 microbeads - by Bioz Stars, 2026-08
97/100 stars
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rantes  (Miltenyi Biotec)
90
Miltenyi Biotec rantes
(A) Freshly isolated NK cells were incubated alone or with CTV-1 cells for 6 hours at 37°C. Supernatants were harvested, and the concentrations of 25 different cytokines and chemokines were determined using a multiplex immunoassay. Bars represent means ± SD of 4 different donors. Post-stimulation cytokine secretion by NK cells was compared with that of cells incubated in medium alone using the paired t- test. Statistical significance is indicated as: *P <0.05; **P <0.01; ***P <0.001. (B) NK cells were incubated with CTV-1 cells at 37°C. Supernatants were harvested at different time points, as indicated, and the concentrations of IFN-γ, <t>TNF-α,</t> <t>MIP-1α,</t> MIP-1β, and <t>RANTES</t> were determined by a multiplex immunoassay. Values represent mean ± SD of 4 different donors.
Rantes, supplied by Miltenyi Biotec, 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/product/105+cells/pmc06594622-42-30-43?v=Miltenyi+Biotec
Average 90 stars, based on 1 article reviews
rantes - by Bioz Stars, 2026-08
90/100 stars
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samples osd1 3 heyman  (Gold Biotechnology Inc)
93
Gold Biotechnology Inc samples osd1 3 heyman
(A) Freshly isolated NK cells were incubated alone or with CTV-1 cells for 6 hours at 37°C. Supernatants were harvested, and the concentrations of 25 different cytokines and chemokines were determined using a multiplex immunoassay. Bars represent means ± SD of 4 different donors. Post-stimulation cytokine secretion by NK cells was compared with that of cells incubated in medium alone using the paired t- test. Statistical significance is indicated as: *P <0.05; **P <0.01; ***P <0.001. (B) NK cells were incubated with CTV-1 cells at 37°C. Supernatants were harvested at different time points, as indicated, and the concentrations of IFN-γ, <t>TNF-α,</t> <t>MIP-1α,</t> MIP-1β, and <t>RANTES</t> were determined by a multiplex immunoassay. Values represent mean ± SD of 4 different donors.
Samples Osd1 3 Heyman, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/105+cells/pm41923635-203-53-49?v=Gold+Biotechnology+Inc
Average 93 stars, based on 1 article reviews
samples osd1 3 heyman - by Bioz Stars, 2026-08
93/100 stars
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anti human cd22 rea340 apc  (Miltenyi Biotec)
92
Miltenyi Biotec anti human cd22 rea340 apc

Anti Human Cd22 Rea340 Apc, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/105+cells/pmc08809252-105-0-5?v=Miltenyi+Biotec
Average 92 stars, based on 1 article reviews
anti human cd22 rea340 apc - by Bioz Stars, 2026-08
92/100 stars
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human epidermal melanocytes  (Cell Applications Inc)
94
Cell Applications Inc human epidermal melanocytes

Human Epidermal Melanocytes, supplied by Cell Applications 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/105+cells/pmc12877985-85-5-8?v=Cell+Applications+Inc
Average 94 stars, based on 1 article reviews
human epidermal melanocytes - by Bioz Stars, 2026-08
94/100 stars
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oadc  (Chem Impex International)
95
Chem Impex International oadc

Oadc, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/105+cells/pm38394154-55-15-53?v=Chem+Impex+International
Average 95 stars, based on 1 article reviews
oadc - by Bioz Stars, 2026-08
95/100 stars
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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
https://www.bioz.com/product/105+cells/pmc08279472-40-3-5?v=StressMarq
Average 93 stars, based on 1 article reviews
gp96 - by Bioz Stars, 2026-08
93/100 stars
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capillary flow cuvette with 2 cross-section  (HELLMA)
90
HELLMA capillary flow cuvette with 2 cross-section
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.
Capillary Flow Cuvette With 2 Cross Section, supplied by HELLMA, 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/product/105+cells/pmc06800439-212-18-20?v=HELLMA
Average 90 stars, based on 1 article reviews
capillary flow cuvette with 2 cross-section - by Bioz Stars, 2026-08
90/100 stars
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quartz cell reference 105.202-qs.10  (HELLMA)
90
HELLMA quartz cell reference 105.202-qs.10
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.
Quartz Cell Reference 105.202 Qs.10, supplied by HELLMA, 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/product/105+cells/pmc08526496-247-12-14?v=HELLMA
Average 90 stars, based on 1 article reviews
quartz cell reference 105.202-qs.10 - by Bioz Stars, 2026-08
90/100 stars
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large-pitch prl 105, single-cell kagome fiber  (KAGOME CO)
90
KAGOME CO large-pitch prl 105, single-cell kagome fiber
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.
Large Pitch Prl 105, Single Cell Kagome Fiber, supplied by KAGOME CO, 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/product/105+cells/10__1103_slash_physrevlett__105__123603-31-8-39?v=KAGOME+CO
Average 90 stars, based on 1 article reviews
large-pitch prl 105, single-cell kagome fiber - by Bioz Stars, 2026-08
90/100 stars
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Image Search Results


(A) Freshly isolated NK cells were incubated alone or with CTV-1 cells for 6 hours at 37°C. Supernatants were harvested, and the concentrations of 25 different cytokines and chemokines were determined using a multiplex immunoassay. Bars represent means ± SD of 4 different donors. Post-stimulation cytokine secretion by NK cells was compared with that of cells incubated in medium alone using the paired t- test. Statistical significance is indicated as: *P <0.05; **P <0.01; ***P <0.001. (B) NK cells were incubated with CTV-1 cells at 37°C. Supernatants were harvested at different time points, as indicated, and the concentrations of IFN-γ, TNF-α, MIP-1α, MIP-1β, and RANTES were determined by a multiplex immunoassay. Values represent mean ± SD of 4 different donors.

Journal: PLoS ONE

Article Title: Tumor- and cytokine-primed human natural killer cells exhibit distinct phenotypic and transcriptional signatures

doi: 10.1371/journal.pone.0218674

Figure Lengend Snippet: (A) Freshly isolated NK cells were incubated alone or with CTV-1 cells for 6 hours at 37°C. Supernatants were harvested, and the concentrations of 25 different cytokines and chemokines were determined using a multiplex immunoassay. Bars represent means ± SD of 4 different donors. Post-stimulation cytokine secretion by NK cells was compared with that of cells incubated in medium alone using the paired t- test. Statistical significance is indicated as: *P <0.05; **P <0.01; ***P <0.001. (B) NK cells were incubated with CTV-1 cells at 37°C. Supernatants were harvested at different time points, as indicated, and the concentrations of IFN-γ, TNF-α, MIP-1α, MIP-1β, and RANTES were determined by a multiplex immunoassay. Values represent mean ± SD of 4 different donors.

Article Snippet: NKp80 (clone 4A4.D10), CD16 (clone VEP3), CD137 (4-1BB; clone 4b4-1), IFN-γ (clone 45–15), TNF-α (clone cA2), MIP-1α (CCL3; clone REA257), MIP-1β (CCL4; clone REA511), MICA/B (clone 6D4), CD155 (clone PV404.19), RANTES (CCL5; clone REA346), and CD244 (2B4; clone REA112) mAbs were purchased from Miltenyi Biotech.

Techniques: Isolation, Incubation, Multiplex Assay

NK cells were incubated in medium alone, with K562, or CTV-1 cells for 6 hours or with IL-2 or IL-12 overnight at 37°C. Supernatants were harvested, and the concentrations of IFN-γ, TNF-α, MIP-1α, MIP-1β, RANTES, IL-R2α, IL-1α, and IL-1β were determined by a multiplex immunoassay. Bars represent means ± SD of 4 different donors. Post-stimulation NK cell cytokine secretion was compared with that in cells incubated in medium alone using the paired t- test. Statistical significance is indicated as: *P <0.05; **P <0.01; ***P <0.001.

Journal: PLoS ONE

Article Title: Tumor- and cytokine-primed human natural killer cells exhibit distinct phenotypic and transcriptional signatures

doi: 10.1371/journal.pone.0218674

Figure Lengend Snippet: NK cells were incubated in medium alone, with K562, or CTV-1 cells for 6 hours or with IL-2 or IL-12 overnight at 37°C. Supernatants were harvested, and the concentrations of IFN-γ, TNF-α, MIP-1α, MIP-1β, RANTES, IL-R2α, IL-1α, and IL-1β were determined by a multiplex immunoassay. Bars represent means ± SD of 4 different donors. Post-stimulation NK cell cytokine secretion was compared with that in cells incubated in medium alone using the paired t- test. Statistical significance is indicated as: *P <0.05; **P <0.01; ***P <0.001.

Article Snippet: NKp80 (clone 4A4.D10), CD16 (clone VEP3), CD137 (4-1BB; clone 4b4-1), IFN-γ (clone 45–15), TNF-α (clone cA2), MIP-1α (CCL3; clone REA257), MIP-1β (CCL4; clone REA511), MICA/B (clone 6D4), CD155 (clone PV404.19), RANTES (CCL5; clone REA346), and CD244 (2B4; clone REA112) mAbs were purchased from Miltenyi Biotech.

Techniques: Incubation, Multiplex Assay

Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet:

Article Snippet: Anti-Human CD22 (REA340) APC , Miltenyi Biotec , 130-120-762; RRID: AB_2752186.

Techniques: Purification, Recombinant, Staining, cDNA Synthesis, Gene Expression, Software, Microscopy

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: