particles (30 to) Search Results


gper1 shgper1  (OriGene)
90
OriGene gper1 shgper1
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Gper1 Shgper1, supplied by OriGene, 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/particles+(30+to)/G+protein+coupled+receptor+30+(GPER1)+Human+shRNA+Lentiviral+Particle/pm32401166-65-19-21
Average 90 stars, based on 1 article reviews
gper1 shgper1 - by Bioz Stars, 2026-09
90/100 stars
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concentrated hcl  (Chem Impex International)
95
Chem Impex International concentrated hcl
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Concentrated Hcl, 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/particles+(30+to)/Hydrochloric+acid/pm31145837-69-9-17
Average 95 stars, based on 1 article reviews
concentrated hcl - by Bioz Stars, 2026-09
95/100 stars
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standard single particle cryo em micrographs  (Gatan Inc)
99
Gatan Inc standard single particle cryo em micrographs
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Standard Single Particle Cryo Em Micrographs, supplied by Gatan Inc, 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/particles+(30+to)/Latitude+S+Software/pmc04775836-41-5-35
Average 99 stars, based on 1 article reviews
standard single particle cryo em micrographs - by Bioz Stars, 2026-09
99/100 stars
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super-fine silica treated with polymerizable organic silane compounds (no. 30, number-average particle size 4 μm  (Mizusawa Industrial Chemicals Ltd)
90
Mizusawa Industrial Chemicals Ltd super-fine silica treated with polymerizable organic silane compounds (no. 30, number-average particle size 4 μm
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Super Fine Silica Treated With Polymerizable Organic Silane Compounds (No. 30, Number Average Particle Size 4 μm, supplied by Mizusawa Industrial Chemicals Ltd, 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/particles+(30+to)/super+fine+silica+treated+with+polymerizable+organic+silane+compounds++no++30++number+average+particle+size+4+%CE%BCm/us07238642-525-58-69
Average 90 stars, based on 1 article reviews
super-fine silica treated with polymerizable organic silane compounds (no. 30, number-average particle size 4 μm - by Bioz Stars, 2026-09
90/100 stars
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diamond/30 μm particle size egg-shaped bur  (Komet GmbH)
90
Komet GmbH diamond/30 μm particle size egg-shaped bur
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Diamond/30 μm Particle Size Egg Shaped Bur, supplied by Komet GmbH, 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/particles+(30+to)/diamond+30+%CE%BCm+particle+size+egg+shaped+bur/pmc06786958-667-29-42
Average 90 stars, based on 1 article reviews
diamond/30 μm particle size egg-shaped bur - by Bioz Stars, 2026-09
90/100 stars
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silicon nanoparticles 30-50 average particle size  (NanoAmor Inc)
90
NanoAmor Inc silicon nanoparticles 30-50 average particle size
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Silicon Nanoparticles 30 50 Average Particle Size, supplied by NanoAmor Inc, 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/particles+(30+to)/silicon+nanoparticles+30+50+average+particle+size/us11094933-106-0-11
Average 90 stars, based on 1 article reviews
silicon nanoparticles 30-50 average particle size - by Bioz Stars, 2026-09
90/100 stars
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dynaron® 8600p  (JSR Corporation)
90
JSR Corporation dynaron® 8600p
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Dynaron® 8600p, supplied by JSR 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/product/particles+(30+to)/dynaron+8600p+made+by+jsr++inc+++styrene+content+15+++mfr+30+g+10+min+++average+dispersed+particle+size+about+20+nm+/us11512185-411-62-66
Average 90 stars, based on 1 article reviews
dynaron® 8600p - by Bioz Stars, 2026-09
90/100 stars
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aqueous dispersion (dispersez-200w2, 30 wt% ptfe content, 200–300 nm particle size)  (Polysciences inc)
90
Polysciences inc aqueous dispersion (dispersez-200w2, 30 wt% ptfe content, 200–300 nm particle size)
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Aqueous Dispersion (Dispersez 200w2, 30 Wt% Ptfe Content, 200–300 Nm Particle Size), supplied by Polysciences inc, 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/particles+(30+to)/aqueous+dispersion++dispersez+200w2++30+wt++ptfe+content++200+300+nm+particle+size+/pmc04342516-49-59-49
Average 90 stars, based on 1 article reviews
aqueous dispersion (dispersez-200w2, 30 wt% ptfe content, 200–300 nm particle size) - by Bioz Stars, 2026-09
90/100 stars
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eurospher ii column (pore size: 100 å, particle size: 5 µm, length: 250 nm, inner diameter: 30 mm, flow rate: 10 ml/min, detection at = 254 nm)  (KNAUER Wissenschaftliche)
90
KNAUER Wissenschaftliche eurospher ii column (pore size: 100 å, particle size: 5 µm, length: 250 nm, inner diameter: 30 mm, flow rate: 10 ml/min, detection at = 254 nm)
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Eurospher Ii Column (Pore Size: 100 å, Particle Size: 5 µm, Length: 250 Nm, Inner Diameter: 30 Mm, Flow Rate: 10 Ml/Min, Detection At = 254 Nm), supplied by KNAUER Wissenschaftliche, 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/particles+(30+to)/eurospher+ii+column++pore+size++100+%C3%A5++particle+size++5+%C2%B5m++length++250+nm++inner+diameter++30+mm++flow+rate++10+ml+min++detection+at++++254+nm+/pmc07202420__OPEN___9___568___s001-27-15-7
Average 90 stars, based on 1 article reviews
eurospher ii column (pore size: 100 å, particle size: 5 µm, length: 250 nm, inner diameter: 30 mm, flow rate: 10 ml/min, detection at = 254 nm) - by Bioz Stars, 2026-09
90/100 stars
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tracer-laden gac particle size 12 × 30 mesh (0.6–1.7)  (Barnebey Sutcliffe Corporation)
90
Barnebey Sutcliffe Corporation tracer-laden gac particle size 12 × 30 mesh (0.6–1.7)
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Tracer Laden Gac Particle Size 12 × 30 Mesh (0.6–1.7), supplied by Barnebey Sutcliffe 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/product/particles+(30+to)/tracer+laden+gac+particle+size+12+%C3%97+30+mesh++0+6+1+7+/pmc03582699-121-9-18
Average 90 stars, based on 1 article reviews
tracer-laden gac particle size 12 × 30 mesh (0.6–1.7) - by Bioz Stars, 2026-09
90/100 stars
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zorbax c-8 guard column (6.0 by 40 mm)  (MAC-MOD Analytical)
90
MAC-MOD Analytical zorbax c-8 guard column (6.0 by 40 mm)
Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on <t>GPER1.</t> (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with <t>shGPER1</t> and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.
Zorbax C 8 Guard Column (6.0 By 40 Mm), supplied by MAC-MOD Analytical, 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/particles+(30+to)/zorbax+sb+c8+column++4+6+%C3%97+30+mm++3+5+%C2%B5m+particles+/10__1128_slash_aac__39__6__1247-91-17-25
Average 90 stars, based on 1 article reviews
zorbax c-8 guard column (6.0 by 40 mm) - by Bioz Stars, 2026-09
90/100 stars
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apc calibration particles (acp-30-5k)  (Spherotech inc)
90
Spherotech inc apc calibration particles (acp-30-5k)
Design and expression of mbaIL6. (A) Schema of the mbaIL6 construct. (B) mbaIL6 and GFP expression in Jurkat cells transduced with either GFP alone (“Control”) or GFP plus mbaIL6, labeled with biotin-conjugated goat anti-human F(ab′)2 and <t>streptavidin-APC.</t> (C) IL-6 binding of control or mbaIL6-transduced Jurkat cells, labeled with IL-6 biotin and streptavidin-APC. Soybean trypsin inhibitor (STI)–biotin was used as a labeling control. (D) Control or mbaIL6-transduced Jurkat cells were labeled with IL-6 biotin and streptavidin-APC; confocal images were captured with a 60× objective lens. (E) Log10 molecules of equivalent APC (MEAPC) and log10 mean fluorescence intensity (MFI) were plotted on a standard curve constructed using <t>allophycocyanin</t> <t>calibration</t> particles (ACP-30-5K). Using the MFI of IL-6–biotin bound to mbaIL6 in the transduced Jurkat cells, the average number of receptors per cell was estimated as follows: log MFI of mbaIL6, 5.27956; log molecules of equivalent APC, 4.321 by intrapolation; yielding 104.321 = 20 941 mbaIL6 receptors per cell. VH, variable heavy chain; VL, variable light chain.
Apc Calibration Particles (Acp 30 5k), supplied by Spherotech inc, 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/particles+(30+to)/apc+calibration+particles++acp+30+5k+/pmc07160280-100-0-4
Average 90 stars, based on 1 article reviews
apc calibration particles (acp-30-5k) - by Bioz Stars, 2026-09
90/100 stars
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Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on GPER1. (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.

Journal: Autophagy

Article Title: Stabilization of MORC2 by estrogen and antiestrogens through GPER1- PRKACA-CMA pathway contributes to estrogen-induced proliferation and endocrine resistance of breast cancer cells.

doi: 10.1080/15548627.2019.1659609

Figure Lengend Snippet: Figure 2. Stabilization of MORC2 by E2 and ESR1 antagonists depends on GPER1. (A-B) SK-BR-3 cells were treated with or without G1 for 24 h at the indicated concentrations (left) or 1 μM of G1 for the indicated times (right) and then subjected to immunoblotting (A) or qPCR (B) analysis. (C-D) SK-BR-3 cells were treated with or without G36 for 24 h at the indicated concentrations (left) or 10 μM of G36 for the indicated times (right) and then subjected to immunoblotting (C) or qPCR (D) analysis. (E-G) SK-BR-3 cells were pretreated with or without 10 nM E2 (E), 10 nM 4-OHT (F), or 10 nM FUL (G) for 1 h and then incubated with 10 μM of G36 for another 24 h. Lysates were collected for immunoblotting analysis. (H) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (Origene) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting. (I-J) SK-BR-3 cells stably expressing shNC and shGPER1 #4 were treated with or without 10 nM E2 (I), 10 nM 4-OHT, or 10 nM FUL (J) for 24 h and then subjected to immunoblotting analysis. (K-M) SK-BR-3 cells were infected with shGPER1 and shNC expression vectors (GenePharma) and selected with 2 μg/ml puromycin for 1 week. The efficacy of shGPER1-mediated knockdown of GPER1 was verified by immunoblotting (K). SK-BR-3 cells stably expressing shNC and shGPER1 #C were treated with or without 10 nM E2 (L), 10 nM 4-OHT, or 10 nM FUL (M) for 24 h and then subjected to immunoblotting analysis.

Article Snippet: Second, we determined whether knockdown of endogenous GPER1 gene in SK-BR-3 cells using specific short hairpin RNAs (shRNAs) targeting GPER1 (shGPER1) (OriGene) could compromiseMORC2 expression induction by E2 and ESR1 MCF-7 0 3 6 12 24 B ESR1 E2 (h) CCND1 MORC2 VCL ESR1 MORC2 VCL C D FUL (h) ESR1 MORC2 VCL SK-BR-3 0 3 6 12 24 MDA-MB-468 0 3 6 12 24 MCF-7 0 3 6 12 24 SK-BR-3 0 3 6 12 24 MDA-MB-468 0 3 6 12 24 MCF-7 0 3 6 12 24 SK-BR-3 0 3 6 12 24 MDA-MB-468 0 3 6 12 24 ESR1 PGR ERBB2 VCL GPER1 A M C F7 SK -B R -3 M D A -M B46 8 E + - - - - + - - - - + - - - - - 3 24 - - - 3 24 - - - 3 24 - - - - - 3 24 - - - 3 24 - - - 3 24 Vehicle 4-OHT (h) FUL (h) G Vehicle E2 Vehicle 0 1.5 3 6 0 1.5 3 6 4-OHT Vehicle 0 1.5 3 6 0 1.5 3 6 FUL 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 M O R C 2/ V C L CHX (h) Vehicle FUL Vehicle 4-OHT Vehicle E2 MORC2 VCL CHX (h) MORC2 VCL 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 Vehicle E2 Vehicle 4-OHT Vehicle FUL M O R C 2/ V C L CHX (h) M C F7 SK -B R -3 H I MCF-7 SK-BR-3 0 1.5 3 6 0 1.5 3 6 4-OHT (h) 1.0 0.9 1.0 1.3 1.9 1.0 3.2 3.4 3.7 3.5 1.0 1.5 1.4 1.8 2.0 1.0 1.5 2.0 2.7 2.7 1.0 1.6 1.7 1.8 2.3 1.0 3.2 3.5 3.4 3.7 1.0 1.0 1.4 1.3 1.8 1.0 1.0 3.4 3.3 3.2 1.0 2.3 6.4 7.2 7.7 ** *** ** **** ** ** **** ** ****** ** ** * MCF-7 SK-BR-3 MDA-MB-468 R el at iv e M O R C 2 m R N A le ve ls 0 0.5 1 1.5 + - - - - + - - - - + - - - - - 3 6 1224 - 3 6 1224 - 3 6 1224 Vehicle E2 (h) 0 1 2 R el at iv e M O R C 2 m R N A le ve lsF MCF-7 SK-BR-3 MDA-MB-468 75 100 150 50 100 150 75 37 100 150 75 100 150 75 100 150 100 150 100 kDa kDa kDa kDa kDa Figure 1.

Techniques: Western Blot, Incubation, Infection, Expressing, Knockdown, Stable Transfection

Figure 3. Activation of PRKACA by GPER1 enhances MORC2 protein stability. (A) SK-BR-3 cells were pretreated with or without 10 μM AG1478, 10 μM LY294002, 10 μM U0126, and 10 μM H89 for 1 h and then incubated with 100 ng/ml EGF for another 30 min. Lysates were subjected to immunoblotting analysis. (B-D) SK-BR-3 cells were pretreated with or without 10 μM AG1478, 10 μM LY294002, 10 μM U0126, and 10 μM H89 for 1 h and then incubated with 10 nM E2 (B), 10 nM 4-OHT (C), or 10 nM FUL (D) for another 24 h. Cells were harvested for immunoblotting analysis. (E-F) SK-BR-3 cells were treated with or without 10 μM forskolin for the indicated times and then subjected to immunoblotting (E) and qPCR (F) analysis. (G-H) SK-BR-3 cells were pretreated with or without 10 μM H89 for 1 h and then treated with or without 10 μM forskolin for another 24 h. Cells were harvested for immunoblotting (G) and qPCR (H) analysis. (I) SK-BR-3 cells were pretreated with or without 10 μM forskolin for 1 h and then incubated with 100 μg/ml of CHX and then analyzed by immunoblotting (upper panel). Relative expression levels of MORC2 are shown in lower panel. (J) SK-BR-3 cells stably expressing shNC and shPRKACA #5 were treated with 100 μg/ml of CHX for the indicated times and then analyzed by immunoblotting (upper panel). Relative expression levels of MORC2 are shown in lower panel. **, p < .01.

Journal: Autophagy

Article Title: Stabilization of MORC2 by estrogen and antiestrogens through GPER1- PRKACA-CMA pathway contributes to estrogen-induced proliferation and endocrine resistance of breast cancer cells.

doi: 10.1080/15548627.2019.1659609

Figure Lengend Snippet: Figure 3. Activation of PRKACA by GPER1 enhances MORC2 protein stability. (A) SK-BR-3 cells were pretreated with or without 10 μM AG1478, 10 μM LY294002, 10 μM U0126, and 10 μM H89 for 1 h and then incubated with 100 ng/ml EGF for another 30 min. Lysates were subjected to immunoblotting analysis. (B-D) SK-BR-3 cells were pretreated with or without 10 μM AG1478, 10 μM LY294002, 10 μM U0126, and 10 μM H89 for 1 h and then incubated with 10 nM E2 (B), 10 nM 4-OHT (C), or 10 nM FUL (D) for another 24 h. Cells were harvested for immunoblotting analysis. (E-F) SK-BR-3 cells were treated with or without 10 μM forskolin for the indicated times and then subjected to immunoblotting (E) and qPCR (F) analysis. (G-H) SK-BR-3 cells were pretreated with or without 10 μM H89 for 1 h and then treated with or without 10 μM forskolin for another 24 h. Cells were harvested for immunoblotting (G) and qPCR (H) analysis. (I) SK-BR-3 cells were pretreated with or without 10 μM forskolin for 1 h and then incubated with 100 μg/ml of CHX and then analyzed by immunoblotting (upper panel). Relative expression levels of MORC2 are shown in lower panel. (J) SK-BR-3 cells stably expressing shNC and shPRKACA #5 were treated with 100 μg/ml of CHX for the indicated times and then analyzed by immunoblotting (upper panel). Relative expression levels of MORC2 are shown in lower panel. **, p < .01.

Article Snippet: Second, we determined whether knockdown of endogenous GPER1 gene in SK-BR-3 cells using specific short hairpin RNAs (shRNAs) targeting GPER1 (shGPER1) (OriGene) could compromiseMORC2 expression induction by E2 and ESR1 MCF-7 0 3 6 12 24 B ESR1 E2 (h) CCND1 MORC2 VCL ESR1 MORC2 VCL C D FUL (h) ESR1 MORC2 VCL SK-BR-3 0 3 6 12 24 MDA-MB-468 0 3 6 12 24 MCF-7 0 3 6 12 24 SK-BR-3 0 3 6 12 24 MDA-MB-468 0 3 6 12 24 MCF-7 0 3 6 12 24 SK-BR-3 0 3 6 12 24 MDA-MB-468 0 3 6 12 24 ESR1 PGR ERBB2 VCL GPER1 A M C F7 SK -B R -3 M D A -M B46 8 E + - - - - + - - - - + - - - - - 3 24 - - - 3 24 - - - 3 24 - - - - - 3 24 - - - 3 24 - - - 3 24 Vehicle 4-OHT (h) FUL (h) G Vehicle E2 Vehicle 0 1.5 3 6 0 1.5 3 6 4-OHT Vehicle 0 1.5 3 6 0 1.5 3 6 FUL 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 M O R C 2/ V C L CHX (h) Vehicle FUL Vehicle 4-OHT Vehicle E2 MORC2 VCL CHX (h) MORC2 VCL 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 Vehicle E2 Vehicle 4-OHT Vehicle FUL M O R C 2/ V C L CHX (h) M C F7 SK -B R -3 H I MCF-7 SK-BR-3 0 1.5 3 6 0 1.5 3 6 4-OHT (h) 1.0 0.9 1.0 1.3 1.9 1.0 3.2 3.4 3.7 3.5 1.0 1.5 1.4 1.8 2.0 1.0 1.5 2.0 2.7 2.7 1.0 1.6 1.7 1.8 2.3 1.0 3.2 3.5 3.4 3.7 1.0 1.0 1.4 1.3 1.8 1.0 1.0 3.4 3.3 3.2 1.0 2.3 6.4 7.2 7.7 ** *** ** **** ** ** **** ** ****** ** ** * MCF-7 SK-BR-3 MDA-MB-468 R el at iv e M O R C 2 m R N A le ve ls 0 0.5 1 1.5 + - - - - + - - - - + - - - - - 3 6 1224 - 3 6 1224 - 3 6 1224 Vehicle E2 (h) 0 1 2 R el at iv e M O R C 2 m R N A le ve lsF MCF-7 SK-BR-3 MDA-MB-468 75 100 150 50 100 150 75 37 100 150 75 100 150 75 100 150 100 150 100 kDa kDa kDa kDa kDa Figure 1.

Techniques: Activation Assay, Incubation, Western Blot, Expressing, Stable Transfection

Figure 9. The proposed working model. E2, TAM, FUL, and G1 stabilize MORC2 in a GPER1 dependent manner. Activated PRKACA kinase by GPER1 phosphorylates MORC2 at T582, which protects MORC2 from lysosomal degradation through blocking its interaction with HSPA8 and LAMP2A. Stabilized MORC2 exerts oncogenic functions to promote E2-induced cell proliferation and decrease cellular sensitivity to antiestrogens.

Journal: Autophagy

Article Title: Stabilization of MORC2 by estrogen and antiestrogens through GPER1- PRKACA-CMA pathway contributes to estrogen-induced proliferation and endocrine resistance of breast cancer cells.

doi: 10.1080/15548627.2019.1659609

Figure Lengend Snippet: Figure 9. The proposed working model. E2, TAM, FUL, and G1 stabilize MORC2 in a GPER1 dependent manner. Activated PRKACA kinase by GPER1 phosphorylates MORC2 at T582, which protects MORC2 from lysosomal degradation through blocking its interaction with HSPA8 and LAMP2A. Stabilized MORC2 exerts oncogenic functions to promote E2-induced cell proliferation and decrease cellular sensitivity to antiestrogens.

Article Snippet: Second, we determined whether knockdown of endogenous GPER1 gene in SK-BR-3 cells using specific short hairpin RNAs (shRNAs) targeting GPER1 (shGPER1) (OriGene) could compromiseMORC2 expression induction by E2 and ESR1 MCF-7 0 3 6 12 24 B ESR1 E2 (h) CCND1 MORC2 VCL ESR1 MORC2 VCL C D FUL (h) ESR1 MORC2 VCL SK-BR-3 0 3 6 12 24 MDA-MB-468 0 3 6 12 24 MCF-7 0 3 6 12 24 SK-BR-3 0 3 6 12 24 MDA-MB-468 0 3 6 12 24 MCF-7 0 3 6 12 24 SK-BR-3 0 3 6 12 24 MDA-MB-468 0 3 6 12 24 ESR1 PGR ERBB2 VCL GPER1 A M C F7 SK -B R -3 M D A -M B46 8 E + - - - - + - - - - + - - - - - 3 24 - - - 3 24 - - - 3 24 - - - - - 3 24 - - - 3 24 - - - 3 24 Vehicle 4-OHT (h) FUL (h) G Vehicle E2 Vehicle 0 1.5 3 6 0 1.5 3 6 4-OHT Vehicle 0 1.5 3 6 0 1.5 3 6 FUL 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 M O R C 2/ V C L CHX (h) Vehicle FUL Vehicle 4-OHT Vehicle E2 MORC2 VCL CHX (h) MORC2 VCL 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 0 0.5 1 1.5 0 1.5 3 6 Vehicle E2 Vehicle 4-OHT Vehicle FUL M O R C 2/ V C L CHX (h) M C F7 SK -B R -3 H I MCF-7 SK-BR-3 0 1.5 3 6 0 1.5 3 6 4-OHT (h) 1.0 0.9 1.0 1.3 1.9 1.0 3.2 3.4 3.7 3.5 1.0 1.5 1.4 1.8 2.0 1.0 1.5 2.0 2.7 2.7 1.0 1.6 1.7 1.8 2.3 1.0 3.2 3.5 3.4 3.7 1.0 1.0 1.4 1.3 1.8 1.0 1.0 3.4 3.3 3.2 1.0 2.3 6.4 7.2 7.7 ** *** ** **** ** ** **** ** ****** ** ** * MCF-7 SK-BR-3 MDA-MB-468 R el at iv e M O R C 2 m R N A le ve ls 0 0.5 1 1.5 + - - - - + - - - - + - - - - - 3 6 1224 - 3 6 1224 - 3 6 1224 Vehicle E2 (h) 0 1 2 R el at iv e M O R C 2 m R N A le ve lsF MCF-7 SK-BR-3 MDA-MB-468 75 100 150 50 100 150 75 37 100 150 75 100 150 75 100 150 100 150 100 kDa kDa kDa kDa kDa Figure 1.

Techniques: Blocking Assay

Design and expression of mbaIL6. (A) Schema of the mbaIL6 construct. (B) mbaIL6 and GFP expression in Jurkat cells transduced with either GFP alone (“Control”) or GFP plus mbaIL6, labeled with biotin-conjugated goat anti-human F(ab′)2 and streptavidin-APC. (C) IL-6 binding of control or mbaIL6-transduced Jurkat cells, labeled with IL-6 biotin and streptavidin-APC. Soybean trypsin inhibitor (STI)–biotin was used as a labeling control. (D) Control or mbaIL6-transduced Jurkat cells were labeled with IL-6 biotin and streptavidin-APC; confocal images were captured with a 60× objective lens. (E) Log10 molecules of equivalent APC (MEAPC) and log10 mean fluorescence intensity (MFI) were plotted on a standard curve constructed using allophycocyanin calibration particles (ACP-30-5K). Using the MFI of IL-6–biotin bound to mbaIL6 in the transduced Jurkat cells, the average number of receptors per cell was estimated as follows: log MFI of mbaIL6, 5.27956; log molecules of equivalent APC, 4.321 by intrapolation; yielding 104.321 = 20 941 mbaIL6 receptors per cell. VH, variable heavy chain; VL, variable light chain.

Journal: Blood Advances

Article Title: Chimeric antigen receptor–T cells with cytokine neutralizing capacity

doi: 10.1182/bloodadvances.2019001287

Figure Lengend Snippet: Design and expression of mbaIL6. (A) Schema of the mbaIL6 construct. (B) mbaIL6 and GFP expression in Jurkat cells transduced with either GFP alone (“Control”) or GFP plus mbaIL6, labeled with biotin-conjugated goat anti-human F(ab′)2 and streptavidin-APC. (C) IL-6 binding of control or mbaIL6-transduced Jurkat cells, labeled with IL-6 biotin and streptavidin-APC. Soybean trypsin inhibitor (STI)–biotin was used as a labeling control. (D) Control or mbaIL6-transduced Jurkat cells were labeled with IL-6 biotin and streptavidin-APC; confocal images were captured with a 60× objective lens. (E) Log10 molecules of equivalent APC (MEAPC) and log10 mean fluorescence intensity (MFI) were plotted on a standard curve constructed using allophycocyanin calibration particles (ACP-30-5K). Using the MFI of IL-6–biotin bound to mbaIL6 in the transduced Jurkat cells, the average number of receptors per cell was estimated as follows: log MFI of mbaIL6, 5.27956; log molecules of equivalent APC, 4.321 by intrapolation; yielding 104.321 = 20 941 mbaIL6 receptors per cell. VH, variable heavy chain; VL, variable light chain.

Article Snippet: APC calibration particles (ACP-30-5K) (Spherotech, Lake Forest, IL) were used to calculate mbaIL6 receptor number per cell.

Techniques: Expressing, Construct, Transduction, Labeling, Binding Assay, Fluorescence

Functional consequences of IL-6 neutralization with mbaIL6-T cells. (A) mbaIL6 and GFP expression in peripheral blood T cells transduced with either GFP alone (“Control”) or GFP plus mbaIL6, after labeling with biotin-conjugated goat anti-human F(ab′)2 antibody and streptavidin-APC. (B) IL-6 binding to Control or mbaIL6-transduced peripheral blood T cells, labeled with IL-6 biotin and streptavidin-APC. (C) Cell marker profile of Control or mbaIL6-transduced T cells from 3 donors. Mean (±SD) of percent T cells expressing each marker is shown. (D) Control or mbaIL6-transduced T lymphocytes at the indicated concentrations were cultured for 2 hours with 1 ng/mL human IL-6; IL-6 in the supernatant was measured by using ELISA. Mean (±SD; n = 3) is shown. ***P < .001. (E) DS-1-mCherry cells were cocultured with Control or mbaIL6-transduced T cells at a 1:1 ratio, with IL-6 (0.5 ng/mL). DS-1 proliferation was quantitated by using the IncuCyte Live Imaging System; shown are mean (±SD) of red calibrated units (RCU) × μm2/well in triplicate measurements. *P = .02; **P < .01 for data at 120 hours. (F) Control or mbaIL6-transduced T lymphocytes at the indicated concentration were cultured for 2 hours with 1 ng/mL IL-6. THP-1 cells were then exposed to either 1 ng/mL IL-6 or to the supernatant of the lymphocyte cultures for 15 minutes at 37°C. Flow cytometry histograms show labeling of THP-1 cells with anti-STAT3 pY705; the graph on the right shows the decrease in pSTAT3 relative to that of THP-1 cells exposed for 15 minutes to 1 ng/mL IL-6. (G) THP-1 cells were exposed to IL-6 for 15 minutes, after 30-minute incubation with the indicated concentrations of tocilizumab. Cells were then labeled with anti-STAT3 pY705 and analyzed as in panel F.

Journal: Blood Advances

Article Title: Chimeric antigen receptor–T cells with cytokine neutralizing capacity

doi: 10.1182/bloodadvances.2019001287

Figure Lengend Snippet: Functional consequences of IL-6 neutralization with mbaIL6-T cells. (A) mbaIL6 and GFP expression in peripheral blood T cells transduced with either GFP alone (“Control”) or GFP plus mbaIL6, after labeling with biotin-conjugated goat anti-human F(ab′)2 antibody and streptavidin-APC. (B) IL-6 binding to Control or mbaIL6-transduced peripheral blood T cells, labeled with IL-6 biotin and streptavidin-APC. (C) Cell marker profile of Control or mbaIL6-transduced T cells from 3 donors. Mean (±SD) of percent T cells expressing each marker is shown. (D) Control or mbaIL6-transduced T lymphocytes at the indicated concentrations were cultured for 2 hours with 1 ng/mL human IL-6; IL-6 in the supernatant was measured by using ELISA. Mean (±SD; n = 3) is shown. ***P < .001. (E) DS-1-mCherry cells were cocultured with Control or mbaIL6-transduced T cells at a 1:1 ratio, with IL-6 (0.5 ng/mL). DS-1 proliferation was quantitated by using the IncuCyte Live Imaging System; shown are mean (±SD) of red calibrated units (RCU) × μm2/well in triplicate measurements. *P = .02; **P < .01 for data at 120 hours. (F) Control or mbaIL6-transduced T lymphocytes at the indicated concentration were cultured for 2 hours with 1 ng/mL IL-6. THP-1 cells were then exposed to either 1 ng/mL IL-6 or to the supernatant of the lymphocyte cultures for 15 minutes at 37°C. Flow cytometry histograms show labeling of THP-1 cells with anti-STAT3 pY705; the graph on the right shows the decrease in pSTAT3 relative to that of THP-1 cells exposed for 15 minutes to 1 ng/mL IL-6. (G) THP-1 cells were exposed to IL-6 for 15 minutes, after 30-minute incubation with the indicated concentrations of tocilizumab. Cells were then labeled with anti-STAT3 pY705 and analyzed as in panel F.

Article Snippet: APC calibration particles (ACP-30-5K) (Spherotech, Lake Forest, IL) were used to calculate mbaIL6 receptor number per cell.

Techniques: Functional Assay, Neutralization, Expressing, Transduction, Labeling, Binding Assay, Marker, Cell Culture, Enzyme-linked Immunosorbent Assay, Imaging, Concentration Assay, Flow Cytometry, Incubation

Design, expression, and IL-6–neutralizing capacity of a bicistronic construct encoding mbaIL6 and anti-CD19 CAR. (A) Schema of the plasmid encoding both receptors (“DUAL”). (B) mbaIL6 expression in peripheral blood T cells transduced with either GFP alone (“Control”), GFP plus anti-CD19 CAR, mbaIL6, or both; cells were labeled with biotin-conjugated goat anti-human F(ab′)2 antibody and streptavidin-APC, and CD19-myc followed by PE-conjugated anti-myc. Expression of each receptor in relation to GFP is shown in supplemental Figure 4. (C) Aggregate data of mbaIL6 and CAR expression from 6 transductions with T cells from 6 donors. Mean (±SD) is shown. (D) Cell marker profile of CAR or DUAL-transduced peripheral blood T cells. Mean (±SD; n = 3) percent T cells expressing each marker is shown. (E) Proportion of naive (CD45RA+ CCR7+), effector (TE, CD45RA+ CCR7‒), central memory (TCM, CD45RA‒ CCR7+), and effector memory (TEM, CD45RA‒ CCR7‒) phenotypes among T cells transduced with the various constructs (mean of 3 experiments). (F) T lymphocytes transduced as in panels B and C were cultured with 1 ng/mL IL-6. After 2 hours, IL-6 levels in the supernatant were measured by using ELISA. Symbols indicate results of 2 independent experiments.

Journal: Blood Advances

Article Title: Chimeric antigen receptor–T cells with cytokine neutralizing capacity

doi: 10.1182/bloodadvances.2019001287

Figure Lengend Snippet: Design, expression, and IL-6–neutralizing capacity of a bicistronic construct encoding mbaIL6 and anti-CD19 CAR. (A) Schema of the plasmid encoding both receptors (“DUAL”). (B) mbaIL6 expression in peripheral blood T cells transduced with either GFP alone (“Control”), GFP plus anti-CD19 CAR, mbaIL6, or both; cells were labeled with biotin-conjugated goat anti-human F(ab′)2 antibody and streptavidin-APC, and CD19-myc followed by PE-conjugated anti-myc. Expression of each receptor in relation to GFP is shown in supplemental Figure 4. (C) Aggregate data of mbaIL6 and CAR expression from 6 transductions with T cells from 6 donors. Mean (±SD) is shown. (D) Cell marker profile of CAR or DUAL-transduced peripheral blood T cells. Mean (±SD; n = 3) percent T cells expressing each marker is shown. (E) Proportion of naive (CD45RA+ CCR7+), effector (TE, CD45RA+ CCR7‒), central memory (TCM, CD45RA‒ CCR7+), and effector memory (TEM, CD45RA‒ CCR7‒) phenotypes among T cells transduced with the various constructs (mean of 3 experiments). (F) T lymphocytes transduced as in panels B and C were cultured with 1 ng/mL IL-6. After 2 hours, IL-6 levels in the supernatant were measured by using ELISA. Symbols indicate results of 2 independent experiments.

Article Snippet: APC calibration particles (ACP-30-5K) (Spherotech, Lake Forest, IL) were used to calculate mbaIL6 receptor number per cell.

Techniques: Expressing, Construct, Plasmid Preparation, Transduction, Labeling, Marker, Cell Culture, Enzyme-linked Immunosorbent Assay

CAR–T cells expressing mbaIL6 quench IL-6 and exert antileukemia activity in xenograft models. (A) NOD/scid-IL2RGnull mice were injected IV with 0.5 to 1 × 106 Nalm-6-luciferase cells. On day 3, mice were given T cells expressing either anti-CD19 CAR alone (85% CAR expression) or mbaIL6 plus CAR (“DUAL”; 79% CAR expression) (20 × 106/mouse IV); all mice received 20 000 IU IL-2 IP every 2 days. Ventral images from the Xenogen IVIS-200 system after D-luciferin injection are shown (captured with enhanced sensitivity on day 3 to visualize Nalm-6 engraftment; full set of ventral and dorsal images is shown in supplemental Figure 7). (B) Luminescence measurements (photons per second) in the mice. Each point corresponds to a measurement in 1 mouse. (C) Levels of GFP+ CD3+ CAR–T cells in blood 50 days after CAR–T cell injection in a subset of the mice. (D) Kaplan-Meier curves of overall survival for the mice shown in panel A, euthanized when the total bioluminescence signal reached 1 × 1010 photons/second. **P < .01 by log-rank test. (E) T cells expressing either anti–CD19 CAR or anti–CD19 CAR plus mbaIL6 were injected IV in NOD/scid-IL2RGnull mice (2-10 × 106/mouse); 3 days later, 50 ng of human IL-6 was injected IP. After 2 hours, mice were euthanized, and serum was collected by cardiac puncture to measure levels of human IL-6 by using ELISA. Each symbol corresponds to data from 1 mouse; bars show mean (±SD). **P < .01. (F) Mice from the experiments shown in panel E were divided according to the number of T cells that were administered: 2 to 4 × 106 (“low”) and 5 to 10 × 106 (“high”). Values correspond to the percentage of IL-6 that was removed from serum in each mouse, using as a reference the mean value of IL-6 measured in mice that received IL-6 with no prior injection of T cells. *P = .035 for CAR low vs DUAL low, P = .045 from CAR high vs, DUAL low, P = .013 for DUAL low vs DUAL high; ***P < .001. (G) Daudi-luciferase cells were injected IP in NOD/scid-IL2RGnull mice (20 × 106/mouse), followed 3 days later by THP-1 and/or T cells IP (20 × 106 for both cell types). Tumor engraftment was measured by in vivo imaging (supplemental Figure 8). Mice were euthanized 48 hours after THP-1 and/or T-cell injection. Symbols show IL-6 levels measured by using ELISA in peritoneal lavage, according to percentage of tumor reduction. *P = .032 for CAR vs no T cells; P = .046 for CAR vs DUAL. (H) IL-6 binding to T cells from the peritoneal lavage of 4 mice, 2 injected with CAR–T cells and 2 injected with T cells expressing both CAR and mbaIL6. Cells were stained with anti-mouse CD45-PE-Cy7, anti-human CD45-PerCP, anti-human CD3-APC, and anti-human IL-6-PE; the plots show selectively gated mouse CD45–, human CD45+, and human CD3+ cells.

Journal: Blood Advances

Article Title: Chimeric antigen receptor–T cells with cytokine neutralizing capacity

doi: 10.1182/bloodadvances.2019001287

Figure Lengend Snippet: CAR–T cells expressing mbaIL6 quench IL-6 and exert antileukemia activity in xenograft models. (A) NOD/scid-IL2RGnull mice were injected IV with 0.5 to 1 × 106 Nalm-6-luciferase cells. On day 3, mice were given T cells expressing either anti-CD19 CAR alone (85% CAR expression) or mbaIL6 plus CAR (“DUAL”; 79% CAR expression) (20 × 106/mouse IV); all mice received 20 000 IU IL-2 IP every 2 days. Ventral images from the Xenogen IVIS-200 system after D-luciferin injection are shown (captured with enhanced sensitivity on day 3 to visualize Nalm-6 engraftment; full set of ventral and dorsal images is shown in supplemental Figure 7). (B) Luminescence measurements (photons per second) in the mice. Each point corresponds to a measurement in 1 mouse. (C) Levels of GFP+ CD3+ CAR–T cells in blood 50 days after CAR–T cell injection in a subset of the mice. (D) Kaplan-Meier curves of overall survival for the mice shown in panel A, euthanized when the total bioluminescence signal reached 1 × 1010 photons/second. **P < .01 by log-rank test. (E) T cells expressing either anti–CD19 CAR or anti–CD19 CAR plus mbaIL6 were injected IV in NOD/scid-IL2RGnull mice (2-10 × 106/mouse); 3 days later, 50 ng of human IL-6 was injected IP. After 2 hours, mice were euthanized, and serum was collected by cardiac puncture to measure levels of human IL-6 by using ELISA. Each symbol corresponds to data from 1 mouse; bars show mean (±SD). **P < .01. (F) Mice from the experiments shown in panel E were divided according to the number of T cells that were administered: 2 to 4 × 106 (“low”) and 5 to 10 × 106 (“high”). Values correspond to the percentage of IL-6 that was removed from serum in each mouse, using as a reference the mean value of IL-6 measured in mice that received IL-6 with no prior injection of T cells. *P = .035 for CAR low vs DUAL low, P = .045 from CAR high vs, DUAL low, P = .013 for DUAL low vs DUAL high; ***P < .001. (G) Daudi-luciferase cells were injected IP in NOD/scid-IL2RGnull mice (20 × 106/mouse), followed 3 days later by THP-1 and/or T cells IP (20 × 106 for both cell types). Tumor engraftment was measured by in vivo imaging (supplemental Figure 8). Mice were euthanized 48 hours after THP-1 and/or T-cell injection. Symbols show IL-6 levels measured by using ELISA in peritoneal lavage, according to percentage of tumor reduction. *P = .032 for CAR vs no T cells; P = .046 for CAR vs DUAL. (H) IL-6 binding to T cells from the peritoneal lavage of 4 mice, 2 injected with CAR–T cells and 2 injected with T cells expressing both CAR and mbaIL6. Cells were stained with anti-mouse CD45-PE-Cy7, anti-human CD45-PerCP, anti-human CD3-APC, and anti-human IL-6-PE; the plots show selectively gated mouse CD45–, human CD45+, and human CD3+ cells.

Article Snippet: APC calibration particles (ACP-30-5K) (Spherotech, Lake Forest, IL) were used to calculate mbaIL6 receptor number per cell.

Techniques: Expressing, Activity Assay, Injection, Luciferase, Enzyme-linked Immunosorbent Assay, In Vivo Imaging, Binding Assay, Staining