xtractor buffer buffer kits Search Results


streptavidin biotin peroxidase complex method  (Agilent technologies)
99
Agilent technologies streptavidin biotin peroxidase complex method
Streptavidin Biotin Peroxidase Complex Method, supplied by Agilent technologies, 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/result/streptavidin biotin peroxidase complex method/product/Agilent technologies
Average 99 stars, based on 1 article reviews
streptavidin biotin peroxidase complex method - by Bioz Stars, 2026-03
99/100 stars
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cell basal media  (ATCC)
96
ATCC cell basal media
Cell Basal Media, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/cell basal media/product/ATCC
Average 96 stars, based on 1 article reviews
cell basal media - by Bioz Stars, 2026-03
96/100 stars
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tmb elisa buffer kit 900-t00  (PeproTech)
90
PeproTech tmb elisa buffer kit 900-t00
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
Tmb Elisa Buffer Kit 900 T00, supplied by PeproTech, 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/tmb elisa buffer kit 900-t00/product/PeproTech
Average 90 stars, based on 1 article reviews
tmb elisa buffer kit 900-t00 - by Bioz Stars, 2026-03
90/100 stars
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flow cytometry fixation & permeabilization buffer kit i  (Bio-Techne corporation)
94
Bio-Techne corporation flow cytometry fixation & permeabilization buffer kit i
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
Flow Cytometry Fixation & Permeabilization Buffer Kit I, supplied by Bio-Techne corporation, 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/result/flow cytometry fixation & permeabilization buffer kit i/product/Bio-Techne corporation
Average 94 stars, based on 1 article reviews
flow cytometry fixation & permeabilization buffer kit i - by Bioz Stars, 2026-03
94/100 stars
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sequencing kit  (Thermo Fisher)
90
Thermo Fisher sequencing kit
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
Sequencing Kit, supplied by Thermo Fisher, 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/sequencing kit/product/Thermo Fisher
Average 90 stars, based on 1 article reviews
sequencing kit - by Bioz Stars, 2026-03
90/100 stars
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rneasy total rna kit  (Qiagen)
90
Qiagen rneasy total rna kit
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
Rneasy Total Rna Kit, supplied by Qiagen, 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/rneasy total rna kit/product/Qiagen
Average 90 stars, based on 1 article reviews
rneasy total rna kit - by Bioz Stars, 2026-03
90/100 stars
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pbs tween buffer  (Thermo Fisher)
99
Thermo Fisher pbs tween buffer
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
Pbs Tween Buffer, supplied by Thermo Fisher, 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/result/pbs tween buffer/product/Thermo Fisher
Average 99 stars, based on 1 article reviews
pbs tween buffer - by Bioz Stars, 2026-03
99/100 stars
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zyme substrate kit  (Bio-Rad)
99
Bio-Rad zyme substrate kit
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
Zyme Substrate Kit, supplied by Bio-Rad, 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/result/zyme substrate kit/product/Bio-Rad
Average 99 stars, based on 1 article reviews
zyme substrate kit - by Bioz Stars, 2026-03
99/100 stars
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normal goat serum  (Vector Laboratories)
96
Vector Laboratories normal goat serum
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
Normal Goat Serum, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/normal goat serum/product/Vector Laboratories
Average 96 stars, based on 1 article reviews
normal goat serum - by Bioz Stars, 2026-03
96/100 stars
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celltag 700 stain  (LI-COR)
98
LI-COR celltag 700 stain
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
Celltag 700 Stain, supplied by LI-COR, 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/result/celltag 700 stain/product/LI-COR
Average 98 stars, based on 1 article reviews
celltag 700 stain - by Bioz Stars, 2026-03
98/100 stars
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10x solution 2  (Miltenyi Biotec)
94
Miltenyi Biotec 10x solution 2
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
10x Solution 2, supplied by Miltenyi Biotec, 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/result/10x solution 2/product/Miltenyi Biotec
Average 94 stars, based on 1 article reviews
10x solution 2 - by Bioz Stars, 2026-03
94/100 stars
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dig dna labeling and detection kit  (Boehringer Mannheim)
90
Boehringer Mannheim dig dna labeling and detection kit
( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by <t>ELISA.</t> ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).
Dig Dna Labeling And Detection Kit, supplied by Boehringer Mannheim, 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/dig dna labeling and detection kit/product/Boehringer Mannheim
Average 90 stars, based on 1 article reviews
dig dna labeling and detection kit - by Bioz Stars, 2026-03
90/100 stars
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Image Search Results


( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by ELISA. ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).

Journal: bioRxiv

Article Title: VPS34-IN1 inhibits cap-mediated translation and synergizes with STING to drive type-I IFN expression in human plasmacytoid DCs

doi: 10.1101/2024.06.17.599308

Figure Lengend Snippet: ( A ) LC3B lipidation was analyzed by determining the LC3-II/LC3-I ratio for CAL-1 cells treated with VPS34-IN1 for 8 h and bafilomycin for 4 h. LC3-I, nonlipidated form; LC3-II, lipidated form. ( B ) CAL-1 cells treated with VPS34-IN1 were stained with MitoSOX and analyzed by flow cytometry. The mean fluorescence intensity was compared to untreated cells (ctr). ( C ) Immunofluorescence confocal microscopy of VPS34-IN1-treated CAL-1 cells showing EEA1 (red) and LAMP1 (blue) distribution (n = 3). Scale bar, 5 μm or 1 μm in magnified areas. ( D ) Cell viability was determined for cells treated with varying concentration of VPS34-IN1 by propidium iodide exclusion and flow cytometry. ( E - I ) Cells were stimulated with CL307 and VPS34-IN1 or spautin-1. ( E, G, H ) Levels of IFNB1 and TNF mRNA were normalized to housekeeping gene ( GAPDH ) and fold change determined in comparison with untreated cells (ctr). ( F ) Immunoblot for phosphorylated STAT1 (Tyr 701 ). Levels of pSTAT1 relative to β-actin were compared against untreated cells (ctr). ( I ) Secretion of IFN-β and TNF-α was monitored by ELISA. ( A , B , D - I ) Bars show the mean ± SD and each dot shows data from one experiment. ns, non-significant results, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA with Šidák post hoc test (A), paired Student’s t test (B), one-way ANOVA with Dunnett post hoc test (D) and one-way ANOVA with Tukey post hoc test (E-I).

Article Snippet: Human TNF-α was quantified in cell supernatants using a commercially available ELISA kit from PeproTech (900-TM25) and the appropriate commercial buffers (TMB ELISA Buffer Kit, 900-T00, PeproTech).

Techniques: Staining, Flow Cytometry, Fluorescence, Immunofluorescence, Confocal Microscopy, Concentration Assay, Comparison, Western Blot, Enzyme-linked Immunosorbent Assay