saline solution Search Results


hepes buffered saline solution  (PromoCell)
94
PromoCell hepes buffered saline solution
Hepes Buffered Saline Solution, supplied by PromoCell, 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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wildtype col4a3 mice  (Teknova)
94
Teknova wildtype col4a3 mice
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Wildtype Col4a3 Mice, supplied by Teknova, 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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1x with pbs  (Rockland Immunochemicals)
95
Rockland Immunochemicals 1x with pbs
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
1x With Pbs, supplied by Rockland Immunochemicals, 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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saline solution  (Teknova)
94
Teknova saline solution
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Saline Solution, supplied by Teknova, 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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sterile saline solution  (Teknova)
94
Teknova sterile saline solution
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Sterile Saline Solution, supplied by Teknova, 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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sterile saline solution - by Bioz Stars, 2026-06
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glycerol  (Chem Impex International)
95
Chem Impex International glycerol
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Glycerol, 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
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sodium chloride  (Chem Impex International)
95
Chem Impex International sodium chloride
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Sodium Chloride, 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
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Average 95 stars, based on 1 article reviews
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sodium chloride solution  (Teknova)
94
Teknova sodium chloride solution
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Sodium Chloride Solution, supplied by Teknova, 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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Average 94 stars, based on 1 article reviews
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erythrocyte pellet  (Randox)
93
Randox erythrocyte pellet
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Erythrocyte Pellet, supplied by Randox, 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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tris  (Chem Impex International)
95
Chem Impex International tris
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Tris, 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
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phosphate buffered saline  (Antibodies Inc)
90
Antibodies Inc phosphate buffered saline
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Phosphate Buffered Saline, supplied by Antibodies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pbs tds buffer  (Bio-Rad)
99
Bio-Rad pbs tds buffer
(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of <t>Col4a3</t> +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).
Pbs Tds Buffer, 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
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Image Search Results


(a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Modulation of the APOM/S1PR4 Pathway Reduces Podocyte Lipid Overload in Alport Syndrome via Distinct Autophagy and Efflux Mechanisms

doi: 10.1681/ASN.0000000996

Figure Lengend Snippet: (a) Representative Western blot analysis and (b) bar graph quantification of APOM protein expression in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=3 mice). (c) Representative Western blot analysis and (d) bar graph quantification of S1PR4 protein expression in kidney cortices (n=3 mice). (e) Bar graph quantification of sphingosine-1-phosphate (S1P) levels determined by LC-MS analysis in kidney cortices of Col4a3 +/+ and Col4a3 −/− mice (n=5 mice). (f) Representative Western blot analysis and (g) bar graph quantification of APOM protein expression in isolated glomeruli (n=3 mice). (h) Representative Western blot and (i) bar graph quantification of S1PR4 protein expression in glomeruli of Col4a3 +/+ and Col4a3 −/− mice (n=3). (j) Representative Western blot analysis and (k, l) bar graph quantification of APOM and S1PR4 protein expression in tubules of Col4a3 +/+ and Col4a3 −/− mice (n=4 mice). (m) Representative Western blot analysis and (n) bar graph quantification of APOM protein expression in Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (o) Representative Western blot analysis and (p) bar graph quantification of S1PR4 protein expression in of Col4a3 +/+ and Col4a3 −/− podocytes (n=3). (q) Bar graph quantification of secreted APOM protein in culture media of Col4a3 +/+ and Col4a3 −/− podocytes by ELISA (n=4). (r) Caspase 3 activity (fold change in apoptosis) in normal human podocytes treated with S1P, albumin, or S1P/albumin with or without APOM or CYM. (n=3–7). (s) Caspase 3 activity in Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− podocytes treated with APOM or CYM. (n=5–6).

Article Snippet: Four-week-old Col4a3 −/− and wildtype (Col4a3 +/+ ) mice were injected with 0.9% saline solution (Teknova, S5819), APOM-Fc (Sino Biologicals, 13495-H02H), IgG-Fc (Sino Biologicals, 10702-HNAH), or CYM50358 (CYM) (Tocris, 4679).

Techniques: Western Blot, Expressing, Liquid Chromatography with Mass Spectroscopy, Isolation, Enzyme-linked Immunosorbent Assay, Activity Assay

Four-week-old Col4a3 −/− and wildtype (Col4a3 +/+ ) mice were injected with either saline solution (control), recombinant human APOM-Fc (APOM), or the selective S1PR4 antagonist CYM50358 (CYM). APOM was administered intraperitoneally once per weekly at doses of 100 μg, 100 μg, 75 μg, and 50 μg, respectively. CYM was administered intraperitoneally three times per week at 10 mg/kg. Mice were sacrificed at 8 weeks of age. Bar graph quantification of (a) urinary albumin-to-creatinine ratio, (b) BUN, and (c) plasma creatinine (n = 6–8 mice). (d) Periodic Acid-Schiff (PAS) staining (4 μm paraffin sections, 20x magnification) and (e) bar graph quantification of the glomerulosclerotic index scores (n=6–8 mice). (f) Representative images of Picro-Sirius red staining (4 μm paraffin sections, 10x) and (g) bar graph quantification of the fibrotic area calculated as percentage of total cortical area (n = 5–8 mice). (h) Representative confocal images of WT1 (red), synaptopodin (green), and DAPI (blue) staining in glomeruli (63× magnification), and (i) bar graph quantification of WT1+ podocytes per glomerulus (n = 5–8). (j) Transmission electron microscopy (TEM) images of glomerular basement membrane (GBM) and podocyte foot processes (2500× magnification), and (k) bar graph quantification of foot processes per μm GBM (n = 6).

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Modulation of the APOM/S1PR4 Pathway Reduces Podocyte Lipid Overload in Alport Syndrome via Distinct Autophagy and Efflux Mechanisms

doi: 10.1681/ASN.0000000996

Figure Lengend Snippet: Four-week-old Col4a3 −/− and wildtype (Col4a3 +/+ ) mice were injected with either saline solution (control), recombinant human APOM-Fc (APOM), or the selective S1PR4 antagonist CYM50358 (CYM). APOM was administered intraperitoneally once per weekly at doses of 100 μg, 100 μg, 75 μg, and 50 μg, respectively. CYM was administered intraperitoneally three times per week at 10 mg/kg. Mice were sacrificed at 8 weeks of age. Bar graph quantification of (a) urinary albumin-to-creatinine ratio, (b) BUN, and (c) plasma creatinine (n = 6–8 mice). (d) Periodic Acid-Schiff (PAS) staining (4 μm paraffin sections, 20x magnification) and (e) bar graph quantification of the glomerulosclerotic index scores (n=6–8 mice). (f) Representative images of Picro-Sirius red staining (4 μm paraffin sections, 10x) and (g) bar graph quantification of the fibrotic area calculated as percentage of total cortical area (n = 5–8 mice). (h) Representative confocal images of WT1 (red), synaptopodin (green), and DAPI (blue) staining in glomeruli (63× magnification), and (i) bar graph quantification of WT1+ podocytes per glomerulus (n = 5–8). (j) Transmission electron microscopy (TEM) images of glomerular basement membrane (GBM) and podocyte foot processes (2500× magnification), and (k) bar graph quantification of foot processes per μm GBM (n = 6).

Article Snippet: Four-week-old Col4a3 −/− and wildtype (Col4a3 +/+ ) mice were injected with 0.9% saline solution (Teknova, S5819), APOM-Fc (Sino Biologicals, 13495-H02H), IgG-Fc (Sino Biologicals, 10702-HNAH), or CYM50358 (CYM) (Tocris, 4679).

Techniques: Recombinant, Injection, Saline, Control, Clinical Proteomics, Staining, Transmission Assay, Electron Microscopy, Membrane

Bar graph quantification of (a) triglycerides, (b) cholesterol esters, and (c) total cholesterol in kidney cortices of Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− mice treated with APOM or CYM (n=6–8 mice). (d) Representative Oil Red O staining of frozen kidney sections (6μm; 20x magnification) and (e) bar graph quantification (n = 4 mice). (f) Confocal images of Nile Red-stained lipid droplets in podocytes, and (g) bar graph quantification of lipid droplet number in Col4a3 +/+ , Col4a3 −/− , and treated Col4a3 −/− podocytes (n = 4). Nile Red (green), DAPI (blue), CellMask HCS and phalloidin (red); 20× magnification. Bar graph quantification of (h) triglycerides (i) cholesterol esters, and (j) total cholesterol in podocytes from each treatment group (n=3–6).

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Modulation of the APOM/S1PR4 Pathway Reduces Podocyte Lipid Overload in Alport Syndrome via Distinct Autophagy and Efflux Mechanisms

doi: 10.1681/ASN.0000000996

Figure Lengend Snippet: Bar graph quantification of (a) triglycerides, (b) cholesterol esters, and (c) total cholesterol in kidney cortices of Col4a3 +/+ , Col4a3 −/− , and Col4a3 −/− mice treated with APOM or CYM (n=6–8 mice). (d) Representative Oil Red O staining of frozen kidney sections (6μm; 20x magnification) and (e) bar graph quantification (n = 4 mice). (f) Confocal images of Nile Red-stained lipid droplets in podocytes, and (g) bar graph quantification of lipid droplet number in Col4a3 +/+ , Col4a3 −/− , and treated Col4a3 −/− podocytes (n = 4). Nile Red (green), DAPI (blue), CellMask HCS and phalloidin (red); 20× magnification. Bar graph quantification of (h) triglycerides (i) cholesterol esters, and (j) total cholesterol in podocytes from each treatment group (n=3–6).

Article Snippet: Four-week-old Col4a3 −/− and wildtype (Col4a3 +/+ ) mice were injected with 0.9% saline solution (Teknova, S5819), APOM-Fc (Sino Biologicals, 13495-H02H), IgG-Fc (Sino Biologicals, 10702-HNAH), or CYM50358 (CYM) (Tocris, 4679).

Techniques: Recombinant, Staining

(a) Venn diagram showing the number of significantly differentially expressed genes (P < 0.05) between Col4a3 +/+ podocytes and Col4a3 −/− podocytes, and between Col4a3 −/− podocytes and Col4a3 −/− podocytes treated with APOM or CYM, as determined by RNA-seq. (b) Volcano plot of RNA-seq results between Col4a3 +/+ podocytes and Col4a3 −/− podocytes (significant genes in red, P < 0.05). (c) KEGG pathway enrichment analysis of differentially expressed genes between Col4a3 +/+ podocytes and Col4a3 −/− podocytes, ranked by significance (d) Volcano plot of RNA-seq results in the APOM treatment group (significant genes in red, P < 0.05). (e) Volcano plot of RNA-seq results for the CYM treatment group (significant genes are shown as red dots (P < 0.05), non-significant genes as black dots) highlighting the most significantly altered gene. (f) Magnified volcano plot of the CYM treatment group excluding Angptl4, which was disproportionately significant (significant genes are shown as red dots (P < 0.05), non-significant genes as black dots). (g) KEGG pathway enrichment analysis of differentially expressed genes in the CYM treatment group, ranked by significance. (h) Heatmap showing Log2 fold change of significantly altered genes (P < 0.05) in the CYM treatment group involved in lysosomal and autophagy pathways. (i) KEGG pathway enrichment analysis of the shared differentially expressed genes between the Col4a3 +/+ podocytes and Col4a3 −/− podocyte group and the CYM treatment group, ranked by significance.

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Modulation of the APOM/S1PR4 Pathway Reduces Podocyte Lipid Overload in Alport Syndrome via Distinct Autophagy and Efflux Mechanisms

doi: 10.1681/ASN.0000000996

Figure Lengend Snippet: (a) Venn diagram showing the number of significantly differentially expressed genes (P < 0.05) between Col4a3 +/+ podocytes and Col4a3 −/− podocytes, and between Col4a3 −/− podocytes and Col4a3 −/− podocytes treated with APOM or CYM, as determined by RNA-seq. (b) Volcano plot of RNA-seq results between Col4a3 +/+ podocytes and Col4a3 −/− podocytes (significant genes in red, P < 0.05). (c) KEGG pathway enrichment analysis of differentially expressed genes between Col4a3 +/+ podocytes and Col4a3 −/− podocytes, ranked by significance (d) Volcano plot of RNA-seq results in the APOM treatment group (significant genes in red, P < 0.05). (e) Volcano plot of RNA-seq results for the CYM treatment group (significant genes are shown as red dots (P < 0.05), non-significant genes as black dots) highlighting the most significantly altered gene. (f) Magnified volcano plot of the CYM treatment group excluding Angptl4, which was disproportionately significant (significant genes are shown as red dots (P < 0.05), non-significant genes as black dots). (g) KEGG pathway enrichment analysis of differentially expressed genes in the CYM treatment group, ranked by significance. (h) Heatmap showing Log2 fold change of significantly altered genes (P < 0.05) in the CYM treatment group involved in lysosomal and autophagy pathways. (i) KEGG pathway enrichment analysis of the shared differentially expressed genes between the Col4a3 +/+ podocytes and Col4a3 −/− podocyte group and the CYM treatment group, ranked by significance.

Article Snippet: Four-week-old Col4a3 −/− and wildtype (Col4a3 +/+ ) mice were injected with 0.9% saline solution (Teknova, S5819), APOM-Fc (Sino Biologicals, 13495-H02H), IgG-Fc (Sino Biologicals, 10702-HNAH), or CYM50358 (CYM) (Tocris, 4679).

Techniques: Expressing, RNA Sequencing

(a) Representative Western blot analysis and (b) bar graph quantification of the LC3-II/LC-I protein expression ratio in Col4a3 −/− podocytes treated with APOM and CYM (n=3). (c) Representative Western blot analysis and (d) bar graph quantification of p62 protein expression in Col4a3 −/− podocytes following treatment (n=5–6). (e) Representative confocal images and bar graph quantification of (f) lysosome number and the (g) co-localization of lysosomes with lipid droplets. Lysotracker (red), Nile red (green), and DAPI (blue) (20x magnification). (h) Representative Western blot analysis and (i) bar graph quantification of ATGL protein expression in Col4a3 −/− podocytes treated with APOM or CYM (n=3). (j) Bar graph quantification of cholesterol efflux in treated Col4a3 −/− podocytes (n=3–8). (k) Representative Western blot of lysosomal acid lipase (LAL) and LC3-II/LC-I protein expression in kidney cortices of Col4a3 −/− mice treated with vehicle or CYM and (l, m) bar graph quantification (n=3–4 mice). (n) Representative Western blot of LAMP1 protein expression in kidney cortices of Col4a3 −/− mice treated with vehicle or CYM and (o) bar graph quantification (n=3–4 mice).

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Modulation of the APOM/S1PR4 Pathway Reduces Podocyte Lipid Overload in Alport Syndrome via Distinct Autophagy and Efflux Mechanisms

doi: 10.1681/ASN.0000000996

Figure Lengend Snippet: (a) Representative Western blot analysis and (b) bar graph quantification of the LC3-II/LC-I protein expression ratio in Col4a3 −/− podocytes treated with APOM and CYM (n=3). (c) Representative Western blot analysis and (d) bar graph quantification of p62 protein expression in Col4a3 −/− podocytes following treatment (n=5–6). (e) Representative confocal images and bar graph quantification of (f) lysosome number and the (g) co-localization of lysosomes with lipid droplets. Lysotracker (red), Nile red (green), and DAPI (blue) (20x magnification). (h) Representative Western blot analysis and (i) bar graph quantification of ATGL protein expression in Col4a3 −/− podocytes treated with APOM or CYM (n=3). (j) Bar graph quantification of cholesterol efflux in treated Col4a3 −/− podocytes (n=3–8). (k) Representative Western blot of lysosomal acid lipase (LAL) and LC3-II/LC-I protein expression in kidney cortices of Col4a3 −/− mice treated with vehicle or CYM and (l, m) bar graph quantification (n=3–4 mice). (n) Representative Western blot of LAMP1 protein expression in kidney cortices of Col4a3 −/− mice treated with vehicle or CYM and (o) bar graph quantification (n=3–4 mice).

Article Snippet: Four-week-old Col4a3 −/− and wildtype (Col4a3 +/+ ) mice were injected with 0.9% saline solution (Teknova, S5819), APOM-Fc (Sino Biologicals, 13495-H02H), IgG-Fc (Sino Biologicals, 10702-HNAH), or CYM50358 (CYM) (Tocris, 4679).

Techniques: Western Blot, Expressing