dimethyl sulfoxide Search Results


dmso  (MedChemExpress)
98
MedChemExpress dmso
Dmso, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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gfs chemicals co  (GFS Chemicals)
93
GFS Chemicals gfs chemicals co
Gfs Chemicals Co, supplied by GFS Chemicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dimethyl sulfoxide  (Toronto Research Chemicals)
91
Toronto Research Chemicals dimethyl sulfoxide
Dimethyl Sulfoxide, supplied by Toronto Research Chemicals, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dimethyl sulfoxide me2so  (Biosynth Carbosynth)
93
Biosynth Carbosynth dimethyl sulfoxide me2so
FIG. 6. The peptide aldehyde, MG132, inhibits b-secretase cleavage of APPSw in stably transfected CHO cells. A, CHOAPPSw cells were incubated for 6 h with <t>Me2SO</t> vehicle or 20–100 mM MG132. Conditioned medium was collected and analyzed for se- creted APPSwb by ELISA. To control for any interference with the ELISA, MG132 was added to a final concentration of 100 mM after CHOAPPSw cells had conditioned medium for 6 h (white bar labeled 1001). B, an ELISA was also conducted on 10-mg aliquots of CHOAPPSw cell lysates to determine whether MG132 treatment al- tered levels of APPSw in the cells. C, MG132 inhibits secretion of APPSwb in a time-dependent manner. CHOAPPSw cells were incu- bated with either Me2SO vehicle (solid line) or 80 mM MG132 (dashed line) for 0, 0.5, 1, 2, 4, 6, or 8 h. Conditioned medium was collected and the levels of secreted APPSwb determined by ELISA.
Dimethyl Sulfoxide Me2so, supplied by Biosynth Carbosynth, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dmso  (Santa Cruz Biotechnology)
94
Santa Cruz Biotechnology dmso
HO53 treatment induces autophagy in human airway epithelial cells. BCi cells were stimulated for 24 h with different doses of HO53, and 250 nM Rapa was used as a positive control for autophagy induction, and <t>DMSO</t> (final concentration of 0.3%) was used as a solv, all in combination with (+Baf.A1) or without <t>(−Baf.A1)</t> <t>Bafilomycin</t> A1 (100 nM). Treatment of differentiated BCi cells was performed by addition of the compound to the lower chamber of the trans-well insert. Induction of autophagy in the ALI culture ( a ) and undifferentiated BCi ( b ) was evaluated by analysis of LC3B processing on Western blotting. The processing of LC3B-I to LC3B-II was quantified by measurement of the LC3B-II band intensity versus GAPDH loading control and presented as the LC3B-II/GAPDH ratio. Data present average ± SEM from n = 3 independent experiments analyzed by one-way ANOVA with Sidak post hoc test, where * p < 0.05, ** p < 0.01 and ns versus solvent control, & p < 0.05 versus 50 μM HO53, && p < 0.01 versus 12.5 μM HO53. Samples were run in one experiment on separate gels/blots processed in parallel, and full-length blots are presented in Supplementary Figure S7. c Analysis of the autophagy induction by HO53 (75 μM) in ALI-cultured BCi cells by immunostaining of LC3B puncta (green), nuclei (blue), and occludin (red), a tight-junction protein characteristic for the differentiated BCi cells in the ALI culture. The scale bar is 10 μm. Autophagy flux in ALI-cultured BCi was calculated based on number of LC3B+ (positive) puncta using the formula: ( sample + Baf.A1 )/ sample presented as autophagy flux LC3B + puncta. Data present average ± SEM from n = 5 independent experiments analyzed by an unpaired t -test, where * p < 0.05. d TEM analysis of differentiated BCi cells treated with HO53 (75 μM) and Bafilomycin A1 (100 nm) for 24 h. Autophagosomes are indicated by red arrows. TF indicates the trans-well filter/insert; red squares indicate a magnified area with scale bars for images as indicated (from left to right) 2 μm, 1 μm and 200 nm. Representative images of n = 4 trans-well inserts from 2 independent experiments. ns, nonsignificant; solv, solvent control; Rapa, rapamycin; SEM, standard error of mean.
Dmso, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dimethyl sulfoxide  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology dimethyl sulfoxide
HO53 treatment induces autophagy in human airway epithelial cells. BCi cells were stimulated for 24 h with different doses of HO53, and 250 nM Rapa was used as a positive control for autophagy induction, and <t>DMSO</t> (final concentration of 0.3%) was used as a solv, all in combination with (+Baf.A1) or without <t>(−Baf.A1)</t> <t>Bafilomycin</t> A1 (100 nM). Treatment of differentiated BCi cells was performed by addition of the compound to the lower chamber of the trans-well insert. Induction of autophagy in the ALI culture ( a ) and undifferentiated BCi ( b ) was evaluated by analysis of LC3B processing on Western blotting. The processing of LC3B-I to LC3B-II was quantified by measurement of the LC3B-II band intensity versus GAPDH loading control and presented as the LC3B-II/GAPDH ratio. Data present average ± SEM from n = 3 independent experiments analyzed by one-way ANOVA with Sidak post hoc test, where * p < 0.05, ** p < 0.01 and ns versus solvent control, & p < 0.05 versus 50 μM HO53, && p < 0.01 versus 12.5 μM HO53. Samples were run in one experiment on separate gels/blots processed in parallel, and full-length blots are presented in Supplementary Figure S7. c Analysis of the autophagy induction by HO53 (75 μM) in ALI-cultured BCi cells by immunostaining of LC3B puncta (green), nuclei (blue), and occludin (red), a tight-junction protein characteristic for the differentiated BCi cells in the ALI culture. The scale bar is 10 μm. Autophagy flux in ALI-cultured BCi was calculated based on number of LC3B+ (positive) puncta using the formula: ( sample + Baf.A1 )/ sample presented as autophagy flux LC3B + puncta. Data present average ± SEM from n = 5 independent experiments analyzed by an unpaired t -test, where * p < 0.05. d TEM analysis of differentiated BCi cells treated with HO53 (75 μM) and Bafilomycin A1 (100 nm) for 24 h. Autophagosomes are indicated by red arrows. TF indicates the trans-well filter/insert; red squares indicate a magnified area with scale bars for images as indicated (from left to right) 2 μm, 1 μm and 200 nm. Representative images of n = 4 trans-well inserts from 2 independent experiments. ns, nonsignificant; solv, solvent control; Rapa, rapamycin; SEM, standard error of mean.
Dimethyl Sulfoxide, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dimethyl sulfoxide  (MedChemExpress)
99
MedChemExpress dimethyl sulfoxide
HO53 treatment induces autophagy in human airway epithelial cells. BCi cells were stimulated for 24 h with different doses of HO53, and 250 nM Rapa was used as a positive control for autophagy induction, and <t>DMSO</t> (final concentration of 0.3%) was used as a solv, all in combination with (+Baf.A1) or without <t>(−Baf.A1)</t> <t>Bafilomycin</t> A1 (100 nM). Treatment of differentiated BCi cells was performed by addition of the compound to the lower chamber of the trans-well insert. Induction of autophagy in the ALI culture ( a ) and undifferentiated BCi ( b ) was evaluated by analysis of LC3B processing on Western blotting. The processing of LC3B-I to LC3B-II was quantified by measurement of the LC3B-II band intensity versus GAPDH loading control and presented as the LC3B-II/GAPDH ratio. Data present average ± SEM from n = 3 independent experiments analyzed by one-way ANOVA with Sidak post hoc test, where * p < 0.05, ** p < 0.01 and ns versus solvent control, & p < 0.05 versus 50 μM HO53, && p < 0.01 versus 12.5 μM HO53. Samples were run in one experiment on separate gels/blots processed in parallel, and full-length blots are presented in Supplementary Figure S7. c Analysis of the autophagy induction by HO53 (75 μM) in ALI-cultured BCi cells by immunostaining of LC3B puncta (green), nuclei (blue), and occludin (red), a tight-junction protein characteristic for the differentiated BCi cells in the ALI culture. The scale bar is 10 μm. Autophagy flux in ALI-cultured BCi was calculated based on number of LC3B+ (positive) puncta using the formula: ( sample + Baf.A1 )/ sample presented as autophagy flux LC3B + puncta. Data present average ± SEM from n = 5 independent experiments analyzed by an unpaired t -test, where * p < 0.05. d TEM analysis of differentiated BCi cells treated with HO53 (75 μM) and Bafilomycin A1 (100 nm) for 24 h. Autophagosomes are indicated by red arrows. TF indicates the trans-well filter/insert; red squares indicate a magnified area with scale bars for images as indicated (from left to right) 2 μm, 1 μm and 200 nm. Representative images of n = 4 trans-well inserts from 2 independent experiments. ns, nonsignificant; solv, solvent control; Rapa, rapamycin; SEM, standard error of mean.
Dimethyl Sulfoxide, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dimethyl sulfoxide  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc dimethyl sulfoxide
HO53 treatment induces autophagy in human airway epithelial cells. BCi cells were stimulated for 24 h with different doses of HO53, and 250 nM Rapa was used as a positive control for autophagy induction, and <t>DMSO</t> (final concentration of 0.3%) was used as a solv, all in combination with (+Baf.A1) or without <t>(−Baf.A1)</t> <t>Bafilomycin</t> A1 (100 nM). Treatment of differentiated BCi cells was performed by addition of the compound to the lower chamber of the trans-well insert. Induction of autophagy in the ALI culture ( a ) and undifferentiated BCi ( b ) was evaluated by analysis of LC3B processing on Western blotting. The processing of LC3B-I to LC3B-II was quantified by measurement of the LC3B-II band intensity versus GAPDH loading control and presented as the LC3B-II/GAPDH ratio. Data present average ± SEM from n = 3 independent experiments analyzed by one-way ANOVA with Sidak post hoc test, where * p < 0.05, ** p < 0.01 and ns versus solvent control, & p < 0.05 versus 50 μM HO53, && p < 0.01 versus 12.5 μM HO53. Samples were run in one experiment on separate gels/blots processed in parallel, and full-length blots are presented in Supplementary Figure S7. c Analysis of the autophagy induction by HO53 (75 μM) in ALI-cultured BCi cells by immunostaining of LC3B puncta (green), nuclei (blue), and occludin (red), a tight-junction protein characteristic for the differentiated BCi cells in the ALI culture. The scale bar is 10 μm. Autophagy flux in ALI-cultured BCi was calculated based on number of LC3B+ (positive) puncta using the formula: ( sample + Baf.A1 )/ sample presented as autophagy flux LC3B + puncta. Data present average ± SEM from n = 5 independent experiments analyzed by an unpaired t -test, where * p < 0.05. d TEM analysis of differentiated BCi cells treated with HO53 (75 μM) and Bafilomycin A1 (100 nm) for 24 h. Autophagosomes are indicated by red arrows. TF indicates the trans-well filter/insert; red squares indicate a magnified area with scale bars for images as indicated (from left to right) 2 μm, 1 μm and 200 nm. Representative images of n = 4 trans-well inserts from 2 independent experiments. ns, nonsignificant; solv, solvent control; Rapa, rapamycin; SEM, standard error of mean.
Dimethyl Sulfoxide, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dimethyl sulfoxide dmso  (MedChemExpress)
99
MedChemExpress dimethyl sulfoxide dmso
HO53 treatment induces autophagy in human airway epithelial cells. BCi cells were stimulated for 24 h with different doses of HO53, and 250 nM Rapa was used as a positive control for autophagy induction, and <t>DMSO</t> (final concentration of 0.3%) was used as a solv, all in combination with (+Baf.A1) or without <t>(−Baf.A1)</t> <t>Bafilomycin</t> A1 (100 nM). Treatment of differentiated BCi cells was performed by addition of the compound to the lower chamber of the trans-well insert. Induction of autophagy in the ALI culture ( a ) and undifferentiated BCi ( b ) was evaluated by analysis of LC3B processing on Western blotting. The processing of LC3B-I to LC3B-II was quantified by measurement of the LC3B-II band intensity versus GAPDH loading control and presented as the LC3B-II/GAPDH ratio. Data present average ± SEM from n = 3 independent experiments analyzed by one-way ANOVA with Sidak post hoc test, where * p < 0.05, ** p < 0.01 and ns versus solvent control, & p < 0.05 versus 50 μM HO53, && p < 0.01 versus 12.5 μM HO53. Samples were run in one experiment on separate gels/blots processed in parallel, and full-length blots are presented in Supplementary Figure S7. c Analysis of the autophagy induction by HO53 (75 μM) in ALI-cultured BCi cells by immunostaining of LC3B puncta (green), nuclei (blue), and occludin (red), a tight-junction protein characteristic for the differentiated BCi cells in the ALI culture. The scale bar is 10 μm. Autophagy flux in ALI-cultured BCi was calculated based on number of LC3B+ (positive) puncta using the formula: ( sample + Baf.A1 )/ sample presented as autophagy flux LC3B + puncta. Data present average ± SEM from n = 5 independent experiments analyzed by an unpaired t -test, where * p < 0.05. d TEM analysis of differentiated BCi cells treated with HO53 (75 μM) and Bafilomycin A1 (100 nm) for 24 h. Autophagosomes are indicated by red arrows. TF indicates the trans-well filter/insert; red squares indicate a magnified area with scale bars for images as indicated (from left to right) 2 μm, 1 μm and 200 nm. Representative images of n = 4 trans-well inserts from 2 independent experiments. ns, nonsignificant; solv, solvent control; Rapa, rapamycin; SEM, standard error of mean.
Dimethyl Sulfoxide Dmso, supplied by MedChemExpress, 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/dimethyl+sulfoxide/Dimethyl+sulfoxide/pmc12816094-28-8-11
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dimethyl sulfoxide  (Valiant Co Ltd)
97
Valiant Co Ltd dimethyl sulfoxide
HO53 treatment induces autophagy in human airway epithelial cells. BCi cells were stimulated for 24 h with different doses of HO53, and 250 nM Rapa was used as a positive control for autophagy induction, and <t>DMSO</t> (final concentration of 0.3%) was used as a solv, all in combination with (+Baf.A1) or without <t>(−Baf.A1)</t> <t>Bafilomycin</t> A1 (100 nM). Treatment of differentiated BCi cells was performed by addition of the compound to the lower chamber of the trans-well insert. Induction of autophagy in the ALI culture ( a ) and undifferentiated BCi ( b ) was evaluated by analysis of LC3B processing on Western blotting. The processing of LC3B-I to LC3B-II was quantified by measurement of the LC3B-II band intensity versus GAPDH loading control and presented as the LC3B-II/GAPDH ratio. Data present average ± SEM from n = 3 independent experiments analyzed by one-way ANOVA with Sidak post hoc test, where * p < 0.05, ** p < 0.01 and ns versus solvent control, & p < 0.05 versus 50 μM HO53, && p < 0.01 versus 12.5 μM HO53. Samples were run in one experiment on separate gels/blots processed in parallel, and full-length blots are presented in Supplementary Figure S7. c Analysis of the autophagy induction by HO53 (75 μM) in ALI-cultured BCi cells by immunostaining of LC3B puncta (green), nuclei (blue), and occludin (red), a tight-junction protein characteristic for the differentiated BCi cells in the ALI culture. The scale bar is 10 μm. Autophagy flux in ALI-cultured BCi was calculated based on number of LC3B+ (positive) puncta using the formula: ( sample + Baf.A1 )/ sample presented as autophagy flux LC3B + puncta. Data present average ± SEM from n = 5 independent experiments analyzed by an unpaired t -test, where * p < 0.05. d TEM analysis of differentiated BCi cells treated with HO53 (75 μM) and Bafilomycin A1 (100 nm) for 24 h. Autophagosomes are indicated by red arrows. TF indicates the trans-well filter/insert; red squares indicate a magnified area with scale bars for images as indicated (from left to right) 2 μm, 1 μm and 200 nm. Representative images of n = 4 trans-well inserts from 2 independent experiments. ns, nonsignificant; solv, solvent control; Rapa, rapamycin; SEM, standard error of mean.
Dimethyl Sulfoxide, supplied by Valiant Co Ltd, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dimethyl sulfoxide dmso  (Aladdin Scientific Corporation)
94
Aladdin Scientific Corporation dimethyl sulfoxide dmso
HO53 treatment induces autophagy in human airway epithelial cells. BCi cells were stimulated for 24 h with different doses of HO53, and 250 nM Rapa was used as a positive control for autophagy induction, and <t>DMSO</t> (final concentration of 0.3%) was used as a solv, all in combination with (+Baf.A1) or without <t>(−Baf.A1)</t> <t>Bafilomycin</t> A1 (100 nM). Treatment of differentiated BCi cells was performed by addition of the compound to the lower chamber of the trans-well insert. Induction of autophagy in the ALI culture ( a ) and undifferentiated BCi ( b ) was evaluated by analysis of LC3B processing on Western blotting. The processing of LC3B-I to LC3B-II was quantified by measurement of the LC3B-II band intensity versus GAPDH loading control and presented as the LC3B-II/GAPDH ratio. Data present average ± SEM from n = 3 independent experiments analyzed by one-way ANOVA with Sidak post hoc test, where * p < 0.05, ** p < 0.01 and ns versus solvent control, & p < 0.05 versus 50 μM HO53, && p < 0.01 versus 12.5 μM HO53. Samples were run in one experiment on separate gels/blots processed in parallel, and full-length blots are presented in Supplementary Figure S7. c Analysis of the autophagy induction by HO53 (75 μM) in ALI-cultured BCi cells by immunostaining of LC3B puncta (green), nuclei (blue), and occludin (red), a tight-junction protein characteristic for the differentiated BCi cells in the ALI culture. The scale bar is 10 μm. Autophagy flux in ALI-cultured BCi was calculated based on number of LC3B+ (positive) puncta using the formula: ( sample + Baf.A1 )/ sample presented as autophagy flux LC3B + puncta. Data present average ± SEM from n = 5 independent experiments analyzed by an unpaired t -test, where * p < 0.05. d TEM analysis of differentiated BCi cells treated with HO53 (75 μM) and Bafilomycin A1 (100 nm) for 24 h. Autophagosomes are indicated by red arrows. TF indicates the trans-well filter/insert; red squares indicate a magnified area with scale bars for images as indicated (from left to right) 2 μm, 1 μm and 200 nm. Representative images of n = 4 trans-well inserts from 2 independent experiments. ns, nonsignificant; solv, solvent control; Rapa, rapamycin; SEM, standard error of mean.
Dimethyl Sulfoxide Dmso, supplied by Aladdin Scientific Corporation, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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promethazine sulfoxide  (Toronto Research Chemicals)
93
Toronto Research Chemicals promethazine sulfoxide
Chemical structure of (a) <t>promethazine</t> hydrochloride, (b) PMZSO, (c) schisandrin, (d) metronidazole, and (e) bifendate.
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Image Search Results


FIG. 6. The peptide aldehyde, MG132, inhibits b-secretase cleavage of APPSw in stably transfected CHO cells. A, CHOAPPSw cells were incubated for 6 h with Me2SO vehicle or 20–100 mM MG132. Conditioned medium was collected and analyzed for se- creted APPSwb by ELISA. To control for any interference with the ELISA, MG132 was added to a final concentration of 100 mM after CHOAPPSw cells had conditioned medium for 6 h (white bar labeled 1001). B, an ELISA was also conducted on 10-mg aliquots of CHOAPPSw cell lysates to determine whether MG132 treatment al- tered levels of APPSw in the cells. C, MG132 inhibits secretion of APPSwb in a time-dependent manner. CHOAPPSw cells were incu- bated with either Me2SO vehicle (solid line) or 80 mM MG132 (dashed line) for 0, 0.5, 1, 2, 4, 6, or 8 h. Conditioned medium was collected and the levels of secreted APPSwb determined by ELISA.

Journal: Journal of Biological Chemistry

Article Title: The Protease Inhibitor, MG132, Blocks Maturation of the Amyloid Precursor Protein Swedish Mutant Preventing Cleavage by β-Secretase

doi: 10.1074/jbc.m008793200

Figure Lengend Snippet: FIG. 6. The peptide aldehyde, MG132, inhibits b-secretase cleavage of APPSw in stably transfected CHO cells. A, CHOAPPSw cells were incubated for 6 h with Me2SO vehicle or 20–100 mM MG132. Conditioned medium was collected and analyzed for se- creted APPSwb by ELISA. To control for any interference with the ELISA, MG132 was added to a final concentration of 100 mM after CHOAPPSw cells had conditioned medium for 6 h (white bar labeled 1001). B, an ELISA was also conducted on 10-mg aliquots of CHOAPPSw cell lysates to determine whether MG132 treatment al- tered levels of APPSw in the cells. C, MG132 inhibits secretion of APPSwb in a time-dependent manner. CHOAPPSw cells were incu- bated with either Me2SO vehicle (solid line) or 80 mM MG132 (dashed line) for 0, 0.5, 1, 2, 4, 6, or 8 h. Conditioned medium was collected and the levels of secreted APPSwb determined by ELISA.

Article Snippet: The peptide aldehyde, MG132, was dissolved in dimethyl sulfoxide (Me2SO) at a concentration of 10 mM (Peptides International).

Techniques: Stable Transfection, Transfection, Incubation, Enzyme-linked Immunosorbent Assay, Control, Concentration Assay, Labeling

FIG. 7. A comparison of AEBSF and MG132 incubation on in- tracellular production of APPSwb. CHOAPPSw cells were pulsed for 12 min with Tran35S-label. The cells were washed and then chased in complete DMEM containing Me2SO, 80 mM MG132, or 1 mM AEBSF for either 45 or 90 min as described under “Experimental Procedures.” Intracellular APPSwb indicated by an arrow (panel B) and full-length APPSw (panel A) were isolated from cell lysates by sequential immu- noprecipitation with 931 and 945 antisera, respectively. Conditioned medium was isolated and analyzed for secreted APPSwb (panel C) and APPSwa (panel D) by immunoprecipitation using the 931 and 6E10 antibodies, respectively.

Journal: Journal of Biological Chemistry

Article Title: The Protease Inhibitor, MG132, Blocks Maturation of the Amyloid Precursor Protein Swedish Mutant Preventing Cleavage by β-Secretase

doi: 10.1074/jbc.m008793200

Figure Lengend Snippet: FIG. 7. A comparison of AEBSF and MG132 incubation on in- tracellular production of APPSwb. CHOAPPSw cells were pulsed for 12 min with Tran35S-label. The cells were washed and then chased in complete DMEM containing Me2SO, 80 mM MG132, or 1 mM AEBSF for either 45 or 90 min as described under “Experimental Procedures.” Intracellular APPSwb indicated by an arrow (panel B) and full-length APPSw (panel A) were isolated from cell lysates by sequential immu- noprecipitation with 931 and 945 antisera, respectively. Conditioned medium was isolated and analyzed for secreted APPSwb (panel C) and APPSwa (panel D) by immunoprecipitation using the 931 and 6E10 antibodies, respectively.

Article Snippet: The peptide aldehyde, MG132, was dissolved in dimethyl sulfoxide (Me2SO) at a concentration of 10 mM (Peptides International).

Techniques: Comparison, Incubation, Isolation, Immunoprecipitation

FIG. 8. A 2-h preincubation with MG132 blocks intracellular production of APPSwb in CHOAPPSw cells. CHOAPPSw cells were preincubated for 2 h with either Me2SO control or 80 mM MG132. The cells were subsequently starved of methionine and cysteine, pulsed for 12 min with Tran35S-label and chased all in the presence of either Me2SO or 80 mM MG132 for either 45 or 90 min as described under “Experimental Procedures.” Intracellular APPSwb indicated by an ar- row (panel B) and full-length APPSw (panel A) were isolated from cell lysates by sequential immunoprecipitation with 931 and 945 antisera, respectively. Conditioned medium was isolated and analyzed for se- creted APPSwb (panel C) and APPSwa (panel D) by immunoprecipita- tion using the 931 and 6E10 antibodies, respectively.

Journal: Journal of Biological Chemistry

Article Title: The Protease Inhibitor, MG132, Blocks Maturation of the Amyloid Precursor Protein Swedish Mutant Preventing Cleavage by β-Secretase

doi: 10.1074/jbc.m008793200

Figure Lengend Snippet: FIG. 8. A 2-h preincubation with MG132 blocks intracellular production of APPSwb in CHOAPPSw cells. CHOAPPSw cells were preincubated for 2 h with either Me2SO control or 80 mM MG132. The cells were subsequently starved of methionine and cysteine, pulsed for 12 min with Tran35S-label and chased all in the presence of either Me2SO or 80 mM MG132 for either 45 or 90 min as described under “Experimental Procedures.” Intracellular APPSwb indicated by an ar- row (panel B) and full-length APPSw (panel A) were isolated from cell lysates by sequential immunoprecipitation with 931 and 945 antisera, respectively. Conditioned medium was isolated and analyzed for se- creted APPSwb (panel C) and APPSwa (panel D) by immunoprecipita- tion using the 931 and 6E10 antibodies, respectively.

Article Snippet: The peptide aldehyde, MG132, was dissolved in dimethyl sulfoxide (Me2SO) at a concentration of 10 mM (Peptides International).

Techniques: Control, Isolation, Immunoprecipitation

HO53 treatment induces autophagy in human airway epithelial cells. BCi cells were stimulated for 24 h with different doses of HO53, and 250 nM Rapa was used as a positive control for autophagy induction, and DMSO (final concentration of 0.3%) was used as a solv, all in combination with (+Baf.A1) or without (−Baf.A1) Bafilomycin A1 (100 nM). Treatment of differentiated BCi cells was performed by addition of the compound to the lower chamber of the trans-well insert. Induction of autophagy in the ALI culture ( a ) and undifferentiated BCi ( b ) was evaluated by analysis of LC3B processing on Western blotting. The processing of LC3B-I to LC3B-II was quantified by measurement of the LC3B-II band intensity versus GAPDH loading control and presented as the LC3B-II/GAPDH ratio. Data present average ± SEM from n = 3 independent experiments analyzed by one-way ANOVA with Sidak post hoc test, where * p < 0.05, ** p < 0.01 and ns versus solvent control, & p < 0.05 versus 50 μM HO53, && p < 0.01 versus 12.5 μM HO53. Samples were run in one experiment on separate gels/blots processed in parallel, and full-length blots are presented in Supplementary Figure S7. c Analysis of the autophagy induction by HO53 (75 μM) in ALI-cultured BCi cells by immunostaining of LC3B puncta (green), nuclei (blue), and occludin (red), a tight-junction protein characteristic for the differentiated BCi cells in the ALI culture. The scale bar is 10 μm. Autophagy flux in ALI-cultured BCi was calculated based on number of LC3B+ (positive) puncta using the formula: ( sample + Baf.A1 )/ sample presented as autophagy flux LC3B + puncta. Data present average ± SEM from n = 5 independent experiments analyzed by an unpaired t -test, where * p < 0.05. d TEM analysis of differentiated BCi cells treated with HO53 (75 μM) and Bafilomycin A1 (100 nm) for 24 h. Autophagosomes are indicated by red arrows. TF indicates the trans-well filter/insert; red squares indicate a magnified area with scale bars for images as indicated (from left to right) 2 μm, 1 μm and 200 nm. Representative images of n = 4 trans-well inserts from 2 independent experiments. ns, nonsignificant; solv, solvent control; Rapa, rapamycin; SEM, standard error of mean.

Journal: Journal of Innate Immunity

Article Title: The Novel Inducer of Innate Immunity HO53 Stimulates Autophagy in Human Airway Epithelial Cells

doi: 10.1159/000521602

Figure Lengend Snippet: HO53 treatment induces autophagy in human airway epithelial cells. BCi cells were stimulated for 24 h with different doses of HO53, and 250 nM Rapa was used as a positive control for autophagy induction, and DMSO (final concentration of 0.3%) was used as a solv, all in combination with (+Baf.A1) or without (−Baf.A1) Bafilomycin A1 (100 nM). Treatment of differentiated BCi cells was performed by addition of the compound to the lower chamber of the trans-well insert. Induction of autophagy in the ALI culture ( a ) and undifferentiated BCi ( b ) was evaluated by analysis of LC3B processing on Western blotting. The processing of LC3B-I to LC3B-II was quantified by measurement of the LC3B-II band intensity versus GAPDH loading control and presented as the LC3B-II/GAPDH ratio. Data present average ± SEM from n = 3 independent experiments analyzed by one-way ANOVA with Sidak post hoc test, where * p < 0.05, ** p < 0.01 and ns versus solvent control, & p < 0.05 versus 50 μM HO53, && p < 0.01 versus 12.5 μM HO53. Samples were run in one experiment on separate gels/blots processed in parallel, and full-length blots are presented in Supplementary Figure S7. c Analysis of the autophagy induction by HO53 (75 μM) in ALI-cultured BCi cells by immunostaining of LC3B puncta (green), nuclei (blue), and occludin (red), a tight-junction protein characteristic for the differentiated BCi cells in the ALI culture. The scale bar is 10 μm. Autophagy flux in ALI-cultured BCi was calculated based on number of LC3B+ (positive) puncta using the formula: ( sample + Baf.A1 )/ sample presented as autophagy flux LC3B + puncta. Data present average ± SEM from n = 5 independent experiments analyzed by an unpaired t -test, where * p < 0.05. d TEM analysis of differentiated BCi cells treated with HO53 (75 μM) and Bafilomycin A1 (100 nm) for 24 h. Autophagosomes are indicated by red arrows. TF indicates the trans-well filter/insert; red squares indicate a magnified area with scale bars for images as indicated (from left to right) 2 μm, 1 μm and 200 nm. Representative images of n = 4 trans-well inserts from 2 independent experiments. ns, nonsignificant; solv, solvent control; Rapa, rapamycin; SEM, standard error of mean.

Article Snippet: UltroserG (15950-017) was obtained from Pall Life Sciences and DMSO (sc-358801), Bafilomycin A1 (sc-201550), and rapamycin (sc-3504) from Santa Cruz.

Techniques: Positive Control, Concentration Assay, Western Blot, Control, Solvent, Cell Culture, Immunostaining

Chemical structure of (a) promethazine hydrochloride, (b) PMZSO, (c) schisandrin, (d) metronidazole, and (e) bifendate.

Journal: International Journal of Analytical Chemistry

Article Title: Simultaneous Determination of Schisandrin and Promethazine with Its Metabolite in Rat Plasma by HPLC-MS/MS and Its Application to a Pharmacokinetic Study

doi: 10.1155/2019/3497045

Figure Lengend Snippet: Chemical structure of (a) promethazine hydrochloride, (b) PMZSO, (c) schisandrin, (d) metronidazole, and (e) bifendate.

Article Snippet: Promethazine sulfoxide (Batch No. 1-JGC-73-2, purity >98%) was manufactured by Toronto Research Chemicals (Toronto, Canada).

Techniques:

MRM transitions, retention time, and conditions of analytes and internal standards.

Journal: International Journal of Analytical Chemistry

Article Title: Simultaneous Determination of Schisandrin and Promethazine with Its Metabolite in Rat Plasma by HPLC-MS/MS and Its Application to a Pharmacokinetic Study

doi: 10.1155/2019/3497045

Figure Lengend Snippet: MRM transitions, retention time, and conditions of analytes and internal standards.

Article Snippet: Promethazine sulfoxide (Batch No. 1-JGC-73-2, purity >98%) was manufactured by Toronto Research Chemicals (Toronto, Canada).

Techniques:

MS scans for (a) promethazine hydrochloride, (b) PMZSO, (c) schisandrin, (d) metronidazole, and (e) bifendate.

Journal: International Journal of Analytical Chemistry

Article Title: Simultaneous Determination of Schisandrin and Promethazine with Its Metabolite in Rat Plasma by HPLC-MS/MS and Its Application to a Pharmacokinetic Study

doi: 10.1155/2019/3497045

Figure Lengend Snippet: MS scans for (a) promethazine hydrochloride, (b) PMZSO, (c) schisandrin, (d) metronidazole, and (e) bifendate.

Article Snippet: Promethazine sulfoxide (Batch No. 1-JGC-73-2, purity >98%) was manufactured by Toronto Research Chemicals (Toronto, Canada).

Techniques:

Regression equations, linear ranges, and LLOQs of the three compounds.

Journal: International Journal of Analytical Chemistry

Article Title: Simultaneous Determination of Schisandrin and Promethazine with Its Metabolite in Rat Plasma by HPLC-MS/MS and Its Application to a Pharmacokinetic Study

doi: 10.1155/2019/3497045

Figure Lengend Snippet: Regression equations, linear ranges, and LLOQs of the three compounds.

Article Snippet: Promethazine sulfoxide (Batch No. 1-JGC-73-2, purity >98%) was manufactured by Toronto Research Chemicals (Toronto, Canada).

Techniques:

Intra- and interday precision (RSD, %) and accuracy (bias, %) of QC samples.

Journal: International Journal of Analytical Chemistry

Article Title: Simultaneous Determination of Schisandrin and Promethazine with Its Metabolite in Rat Plasma by HPLC-MS/MS and Its Application to a Pharmacokinetic Study

doi: 10.1155/2019/3497045

Figure Lengend Snippet: Intra- and interday precision (RSD, %) and accuracy (bias, %) of QC samples.

Article Snippet: Promethazine sulfoxide (Batch No. 1-JGC-73-2, purity >98%) was manufactured by Toronto Research Chemicals (Toronto, Canada).

Techniques: Concentration Assay

Recovery and matrix effect of the three analytes in rat plasma.

Journal: International Journal of Analytical Chemistry

Article Title: Simultaneous Determination of Schisandrin and Promethazine with Its Metabolite in Rat Plasma by HPLC-MS/MS and Its Application to a Pharmacokinetic Study

doi: 10.1155/2019/3497045

Figure Lengend Snippet: Recovery and matrix effect of the three analytes in rat plasma.

Article Snippet: Promethazine sulfoxide (Batch No. 1-JGC-73-2, purity >98%) was manufactured by Toronto Research Chemicals (Toronto, Canada).

Techniques: Clinical Proteomics, Concentration Assay

Stability of the analytes in rat plasma under different storage conditions.

Journal: International Journal of Analytical Chemistry

Article Title: Simultaneous Determination of Schisandrin and Promethazine with Its Metabolite in Rat Plasma by HPLC-MS/MS and Its Application to a Pharmacokinetic Study

doi: 10.1155/2019/3497045

Figure Lengend Snippet: Stability of the analytes in rat plasma under different storage conditions.

Article Snippet: Promethazine sulfoxide (Batch No. 1-JGC-73-2, purity >98%) was manufactured by Toronto Research Chemicals (Toronto, Canada).

Techniques: Clinical Proteomics, Concentration Assay

Pharmacokinetic parameters of PMZ and PMZSO in rats after oral administration of PMZ with or without S. chinensis ( n = 6-7).

Journal: International Journal of Analytical Chemistry

Article Title: Simultaneous Determination of Schisandrin and Promethazine with Its Metabolite in Rat Plasma by HPLC-MS/MS and Its Application to a Pharmacokinetic Study

doi: 10.1155/2019/3497045

Figure Lengend Snippet: Pharmacokinetic parameters of PMZ and PMZSO in rats after oral administration of PMZ with or without S. chinensis ( n = 6-7).

Article Snippet: Promethazine sulfoxide (Batch No. 1-JGC-73-2, purity >98%) was manufactured by Toronto Research Chemicals (Toronto, Canada).

Techniques: