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  • 94
    Proteintech hsd11b2
    Antibody information for Western blotting verification.
    Hsd11b2, supplied by Proteintech, 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/hsd11b2/product/Proteintech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hsd11b2 - by Bioz Stars, 2024-07
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    94
    Proteintech anti cyp11a1
    Antibody information for Western blotting verification.
    Anti Cyp11a1, supplied by Proteintech, 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/anti cyp11a1/product/Proteintech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cyp11a1 - by Bioz Stars, 2024-07
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    94
    Proteintech anti cyp11a1 rabbit pab
    mRNA expression of <t>CYP11A1</t> in human brain tissues . A , schematic diagram describing the cholesterol side-chain cleavage reaction by CYP11A1. qRT-PCR analyses of CYP11A1 in ( B ) total brain, cerebellum, cortex, parietal lobe, occipital pole, temporal lobe, and spinal cord and ( C ) human testes, adrenal glands, brain, and spinal cord. Data for brain and spinal cord in ( C ) are the same as the total brain and spinal cord data in ( B ), respectively. Gene expression is shown as relative expression to α-tubulin. Data are presented as mean ± SD, N = 3. Each data point represents a replicate of the same RNA sample. ( D – G ) RNAscope in situ hybridization analyses of human cerebellum and cerebral cortex tissue (male, 63-years-old). A chromogenic assay was performed with two targets: CYP11A1 ( red ) and myelin basic protein (MBP; teal ). Each punctate dot represents one molecule of mRNA. CYP11A1-positive cells can be found in the granule layer ( D ), molecular layer and Purkinje layer ( E ) of the cerebellum, as well as gray matter of the cortex ( F ), indicated by black arrows . No CYP11A1-positive cells were observed in white matter ( G ). Little colocalization between MBP and CYP11A1 can be observed. qRT-PCR, quantitative RT-PCR.
    Anti Cyp11a1 Rabbit Pab, supplied by Proteintech, 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/anti cyp11a1 rabbit pab/product/Proteintech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cyp11a1 rabbit pab - by Bioz Stars, 2024-07
    94/100 stars
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    94
    Proteintech rabbit anti cyp11a p450scc
    PS-MPs treatment downregulated the expression of steroidogenic enzymes and StAR in testes. Mice were given drinking water comprising PS-MPs of various sizes for 180 sustained days. A , B The expression of 3β-HSD, 17β-HSD, <t>P450scc,</t> P450c17, and StAR in testes was measured by western blotting. The expression levels were quantified with ImageJ (n = 3). Data are expressed as means ± SD. * P < 0.05, ** P < 0.01 vs. control. # P < 0.05, ## P < 0.01 vs. 100 μg/L group. C , D The expression of StAR in mouse testicular tissues of the control group and 1000 μg/L group was tested by immunofluorescence staining. Testicular tissues were stained with StAR (red) and DAPI (blue), arrows: positive expression (scale bar = 50 µm). The expression levels of StAR were quantified with ImageJ (n = 3). Data are expressed as means ± SD. ** P < 0.01 vs. control. # P < 0.05, *** P < 0.001 vs. control
    Rabbit Anti Cyp11a P450scc, supplied by Proteintech, 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/rabbit anti cyp11a p450scc/product/Proteintech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti cyp11a p450scc - by Bioz Stars, 2024-07
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    94
    Proteintech 1 ap
    PS-MPs treatment downregulated the expression of steroidogenic enzymes and StAR in testes. Mice were given drinking water comprising PS-MPs of various sizes for 180 sustained days. A , B The expression of 3β-HSD, 17β-HSD, <t>P450scc,</t> P450c17, and StAR in testes was measured by western blotting. The expression levels were quantified with ImageJ (n = 3). Data are expressed as means ± SD. * P < 0.05, ** P < 0.01 vs. control. # P < 0.05, ## P < 0.01 vs. 100 μg/L group. C , D The expression of StAR in mouse testicular tissues of the control group and 1000 μg/L group was tested by immunofluorescence staining. Testicular tissues were stained with StAR (red) and DAPI (blue), arrows: positive expression (scale bar = 50 µm). The expression levels of StAR were quantified with ImageJ (n = 3). Data are expressed as means ± SD. ** P < 0.01 vs. control. # P < 0.05, *** P < 0.001 vs. control
    1 Ap, supplied by Proteintech, 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/1 ap/product/Proteintech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    1 ap - by Bioz Stars, 2024-07
    94/100 stars
      Buy from Supplier

    Image Search Results


    Antibody information for Western blotting verification.

    Journal: Scientific Reports

    Article Title: Proteomics analysis reveals the effect of 1α,25(OH) 2 VD 3 -glycosides on development of early testes in piglets

    doi: 10.1038/s41598-021-90676-8

    Figure Lengend Snippet: Antibody information for Western blotting verification.

    Article Snippet: HSD11B2 , Rabbit , Proteintech , 13,363–1-AP , 1:1000 , 40.

    Techniques: Western Blot

    Validation of ten selected proteins in testicular tissue of animals from treatments S1 and S4 by Western blotting. ( A ) The abundance of CYP11A1, CYP19A1, HSD11B2, FDFT1, PEX10, PHYH, HSD17B4, CAT, DHRS4 and INSL3 proteins were analyzed by Western blotting in three replicates. The β-actin was used as an internal reference. ( B ) Comparison and verification of the quantitative results for selected proteins from the Western blotting. The values shown are the means ± SEM (Significant difference: * P < 0.05; ** P < 0.01; *** P < 0.001). S1: control diet; S4: control diet supplemented with 4 μg/kg 1α,25(OH) 2 VD 3. 1α,25(OH) 2 VD 3 was provided as 1α,25(OH) 2 VD 3 -glycosides from plant origin (n = 3 per treatment).

    Journal: Scientific Reports

    Article Title: Proteomics analysis reveals the effect of 1α,25(OH) 2 VD 3 -glycosides on development of early testes in piglets

    doi: 10.1038/s41598-021-90676-8

    Figure Lengend Snippet: Validation of ten selected proteins in testicular tissue of animals from treatments S1 and S4 by Western blotting. ( A ) The abundance of CYP11A1, CYP19A1, HSD11B2, FDFT1, PEX10, PHYH, HSD17B4, CAT, DHRS4 and INSL3 proteins were analyzed by Western blotting in three replicates. The β-actin was used as an internal reference. ( B ) Comparison and verification of the quantitative results for selected proteins from the Western blotting. The values shown are the means ± SEM (Significant difference: * P < 0.05; ** P < 0.01; *** P < 0.001). S1: control diet; S4: control diet supplemented with 4 μg/kg 1α,25(OH) 2 VD 3. 1α,25(OH) 2 VD 3 was provided as 1α,25(OH) 2 VD 3 -glycosides from plant origin (n = 3 per treatment).

    Article Snippet: HSD11B2 , Rabbit , Proteintech , 13,363–1-AP , 1:1000 , 40.

    Techniques: Western Blot

    mRNA expression of CYP11A1 in human brain tissues . A , schematic diagram describing the cholesterol side-chain cleavage reaction by CYP11A1. qRT-PCR analyses of CYP11A1 in ( B ) total brain, cerebellum, cortex, parietal lobe, occipital pole, temporal lobe, and spinal cord and ( C ) human testes, adrenal glands, brain, and spinal cord. Data for brain and spinal cord in ( C ) are the same as the total brain and spinal cord data in ( B ), respectively. Gene expression is shown as relative expression to α-tubulin. Data are presented as mean ± SD, N = 3. Each data point represents a replicate of the same RNA sample. ( D – G ) RNAscope in situ hybridization analyses of human cerebellum and cerebral cortex tissue (male, 63-years-old). A chromogenic assay was performed with two targets: CYP11A1 ( red ) and myelin basic protein (MBP; teal ). Each punctate dot represents one molecule of mRNA. CYP11A1-positive cells can be found in the granule layer ( D ), molecular layer and Purkinje layer ( E ) of the cerebellum, as well as gray matter of the cortex ( F ), indicated by black arrows . No CYP11A1-positive cells were observed in white matter ( G ). Little colocalization between MBP and CYP11A1 can be observed. qRT-PCR, quantitative RT-PCR.

    Journal: The Journal of Biological Chemistry

    Article Title: The neurosteroid pregnenolone is synthesized by a mitochondrial P450 enzyme other than CYP11A1 in human glial cells

    doi: 10.1016/j.jbc.2022.102110

    Figure Lengend Snippet: mRNA expression of CYP11A1 in human brain tissues . A , schematic diagram describing the cholesterol side-chain cleavage reaction by CYP11A1. qRT-PCR analyses of CYP11A1 in ( B ) total brain, cerebellum, cortex, parietal lobe, occipital pole, temporal lobe, and spinal cord and ( C ) human testes, adrenal glands, brain, and spinal cord. Data for brain and spinal cord in ( C ) are the same as the total brain and spinal cord data in ( B ), respectively. Gene expression is shown as relative expression to α-tubulin. Data are presented as mean ± SD, N = 3. Each data point represents a replicate of the same RNA sample. ( D – G ) RNAscope in situ hybridization analyses of human cerebellum and cerebral cortex tissue (male, 63-years-old). A chromogenic assay was performed with two targets: CYP11A1 ( red ) and myelin basic protein (MBP; teal ). Each punctate dot represents one molecule of mRNA. CYP11A1-positive cells can be found in the granule layer ( D ), molecular layer and Purkinje layer ( E ) of the cerebellum, as well as gray matter of the cortex ( F ), indicated by black arrows . No CYP11A1-positive cells were observed in white matter ( G ). Little colocalization between MBP and CYP11A1 can be observed. qRT-PCR, quantitative RT-PCR.

    Article Snippet: The following primary antibodies were used: anti-CYP11A1 rabbit mAb (Cell Signaling Technology; #14217, 1:1000 dilution, antigen: human CYP11A1 N terminus), anti-CYP11A1 rabbit pAb (Proteintech; #13363-1-AP, 1:1000 dilution, antigen: CYP11A1 aa 1–300), anti-CYP11A1 rabbit pAb (Abcam; #ab75497, 1:500 dilution, antigen: human CYP11A1 aa 288–337), anti-CYP11A1 rabbit pAb (Abcam; #ab232763, 1:1000 dilution, antigen: CYP11A1 aa 350–521), anti-GAPDH rabbit mAb (Cell Signaling Technology; #2118, 1:2000 dilution), anti-FDXR rabbit pAb (Proteintech; #15584-1-AP, 1:1000 dilution), anti-ADX rabbit mAb (Abcam; #ab108257, 1:1000 dilution), antibeta actin mouse mAb (Abcam; #ab8226, 1:3000 dilution), anti-CYP reductase rabbit pAb (Abcam; #ab13513, 1:1000 dilution), and anti-Myc-tag rabbit mAb (Cell Signaling Technology; #2278S, 1:1000 dilution).

    Techniques: Expressing, Quantitative RT-PCR, In Situ Hybridization, Chromogenic Assay

    RNA and protein expression of CYP11A1 and its cofactors in glial cells. A – C qRT-PCR analyses of CYP11A1, FDX1, and FDXR in human glial cells, with H295R-S1 as a positive control. Gene expression is shown as relative expression to α-tubulin. Data are presented as mean ± SD, N = 3. Each data point represents total RNA extracted from cells of a different passage for each cell line. D and E representative immunoblots of CYP11A1, FDX1, and FDXR in human glial cells, with MA-10 and H295R-S1 as positive controls. GAPDH was used as a loading control. The 15 μg of total cell lysate was loaded in each lane. No specific bands for CYP11A1 can be observed in glial cells. qRT-PCR, quantitative RT-PCR.

    Journal: The Journal of Biological Chemistry

    Article Title: The neurosteroid pregnenolone is synthesized by a mitochondrial P450 enzyme other than CYP11A1 in human glial cells

    doi: 10.1016/j.jbc.2022.102110

    Figure Lengend Snippet: RNA and protein expression of CYP11A1 and its cofactors in glial cells. A – C qRT-PCR analyses of CYP11A1, FDX1, and FDXR in human glial cells, with H295R-S1 as a positive control. Gene expression is shown as relative expression to α-tubulin. Data are presented as mean ± SD, N = 3. Each data point represents total RNA extracted from cells of a different passage for each cell line. D and E representative immunoblots of CYP11A1, FDX1, and FDXR in human glial cells, with MA-10 and H295R-S1 as positive controls. GAPDH was used as a loading control. The 15 μg of total cell lysate was loaded in each lane. No specific bands for CYP11A1 can be observed in glial cells. qRT-PCR, quantitative RT-PCR.

    Article Snippet: The following primary antibodies were used: anti-CYP11A1 rabbit mAb (Cell Signaling Technology; #14217, 1:1000 dilution, antigen: human CYP11A1 N terminus), anti-CYP11A1 rabbit pAb (Proteintech; #13363-1-AP, 1:1000 dilution, antigen: CYP11A1 aa 1–300), anti-CYP11A1 rabbit pAb (Abcam; #ab75497, 1:500 dilution, antigen: human CYP11A1 aa 288–337), anti-CYP11A1 rabbit pAb (Abcam; #ab232763, 1:1000 dilution, antigen: CYP11A1 aa 350–521), anti-GAPDH rabbit mAb (Cell Signaling Technology; #2118, 1:2000 dilution), anti-FDXR rabbit pAb (Proteintech; #15584-1-AP, 1:1000 dilution), anti-ADX rabbit mAb (Abcam; #ab108257, 1:1000 dilution), antibeta actin mouse mAb (Abcam; #ab8226, 1:3000 dilution), anti-CYP reductase rabbit pAb (Abcam; #ab13513, 1:1000 dilution), and anti-Myc-tag rabbit mAb (Cell Signaling Technology; #2278S, 1:1000 dilution).

    Techniques: Expressing, Quantitative RT-PCR, Positive Control, Western Blot

    Confocal images of H295R-S1, MGM-1, MGM-3, NHA, and HMC3 cells for CYP11A1 expression . Cells stained with anti-CYP11A1 antibody ( green ), with mitochondria labeled with Mitotracker Red and nucleus stained with DAPI ( blue ). Images were taken at 63× magnification. H295R-S1 cells stain very strongly for CYP11A1, which overlaps entirely with mitochondrial staining. Very faint CYP11A1 staining colocalizing with mitochondria can be seen in MGM-1, MGM-3, NHA, and HMC3 cells, with the strongest signal in MGM-3 cells. DAPI, 4′,6-diamidino-2-phenylindole; NHA, normal human astrocyte.

    Journal: The Journal of Biological Chemistry

    Article Title: The neurosteroid pregnenolone is synthesized by a mitochondrial P450 enzyme other than CYP11A1 in human glial cells

    doi: 10.1016/j.jbc.2022.102110

    Figure Lengend Snippet: Confocal images of H295R-S1, MGM-1, MGM-3, NHA, and HMC3 cells for CYP11A1 expression . Cells stained with anti-CYP11A1 antibody ( green ), with mitochondria labeled with Mitotracker Red and nucleus stained with DAPI ( blue ). Images were taken at 63× magnification. H295R-S1 cells stain very strongly for CYP11A1, which overlaps entirely with mitochondrial staining. Very faint CYP11A1 staining colocalizing with mitochondria can be seen in MGM-1, MGM-3, NHA, and HMC3 cells, with the strongest signal in MGM-3 cells. DAPI, 4′,6-diamidino-2-phenylindole; NHA, normal human astrocyte.

    Article Snippet: The following primary antibodies were used: anti-CYP11A1 rabbit mAb (Cell Signaling Technology; #14217, 1:1000 dilution, antigen: human CYP11A1 N terminus), anti-CYP11A1 rabbit pAb (Proteintech; #13363-1-AP, 1:1000 dilution, antigen: CYP11A1 aa 1–300), anti-CYP11A1 rabbit pAb (Abcam; #ab75497, 1:500 dilution, antigen: human CYP11A1 aa 288–337), anti-CYP11A1 rabbit pAb (Abcam; #ab232763, 1:1000 dilution, antigen: CYP11A1 aa 350–521), anti-GAPDH rabbit mAb (Cell Signaling Technology; #2118, 1:2000 dilution), anti-FDXR rabbit pAb (Proteintech; #15584-1-AP, 1:1000 dilution), anti-ADX rabbit mAb (Abcam; #ab108257, 1:1000 dilution), antibeta actin mouse mAb (Abcam; #ab8226, 1:3000 dilution), anti-CYP reductase rabbit pAb (Abcam; #ab13513, 1:1000 dilution), and anti-Myc-tag rabbit mAb (Cell Signaling Technology; #2278S, 1:1000 dilution).

    Techniques: Expressing, Staining, Labeling

    Expression of CYP11A1, FDXR, and POR in MGM-1 cells after treatments that increase pregnenolone synthesis . A – C , qRT-PCR analysis of CYP11A1 expression ( A ) in addition to expression of CYP450 cofactors, FDXR ( B ) and POR ( C ), following treatment of 0.76 mM AMG, 50 μM ketoconazole (KC), or 50 μM 22(R)-hydroxycholesterol (22(R)-HC) for 2 h in MGM-1 cells. Gene expression is shown as relative expression to β-actin. Data are presented as mean ± SD, N = 3. D , representative immunoblot showing protein expression of CYP11A1, FDXR, and POR following the aforementioned treatments. β-Actin was used as a loading control. No significant changes were observed for either RNA or protein expression of CYP11A1, FDXR, and POR following AMG, KC, and 22(R)-HC treatments. AMG, aminoglutethimide; qRT-PCR, quantitative RT-PCR.

    Journal: The Journal of Biological Chemistry

    Article Title: The neurosteroid pregnenolone is synthesized by a mitochondrial P450 enzyme other than CYP11A1 in human glial cells

    doi: 10.1016/j.jbc.2022.102110

    Figure Lengend Snippet: Expression of CYP11A1, FDXR, and POR in MGM-1 cells after treatments that increase pregnenolone synthesis . A – C , qRT-PCR analysis of CYP11A1 expression ( A ) in addition to expression of CYP450 cofactors, FDXR ( B ) and POR ( C ), following treatment of 0.76 mM AMG, 50 μM ketoconazole (KC), or 50 μM 22(R)-hydroxycholesterol (22(R)-HC) for 2 h in MGM-1 cells. Gene expression is shown as relative expression to β-actin. Data are presented as mean ± SD, N = 3. D , representative immunoblot showing protein expression of CYP11A1, FDXR, and POR following the aforementioned treatments. β-Actin was used as a loading control. No significant changes were observed for either RNA or protein expression of CYP11A1, FDXR, and POR following AMG, KC, and 22(R)-HC treatments. AMG, aminoglutethimide; qRT-PCR, quantitative RT-PCR.

    Article Snippet: The following primary antibodies were used: anti-CYP11A1 rabbit mAb (Cell Signaling Technology; #14217, 1:1000 dilution, antigen: human CYP11A1 N terminus), anti-CYP11A1 rabbit pAb (Proteintech; #13363-1-AP, 1:1000 dilution, antigen: CYP11A1 aa 1–300), anti-CYP11A1 rabbit pAb (Abcam; #ab75497, 1:500 dilution, antigen: human CYP11A1 aa 288–337), anti-CYP11A1 rabbit pAb (Abcam; #ab232763, 1:1000 dilution, antigen: CYP11A1 aa 350–521), anti-GAPDH rabbit mAb (Cell Signaling Technology; #2118, 1:2000 dilution), anti-FDXR rabbit pAb (Proteintech; #15584-1-AP, 1:1000 dilution), anti-ADX rabbit mAb (Abcam; #ab108257, 1:1000 dilution), antibeta actin mouse mAb (Abcam; #ab8226, 1:3000 dilution), anti-CYP reductase rabbit pAb (Abcam; #ab13513, 1:1000 dilution), and anti-Myc-tag rabbit mAb (Cell Signaling Technology; #2278S, 1:1000 dilution).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    RNA Expression of CYP11A1 variants in the human CNS and peripheral tissues and cells . qRT-PCR analyses of total CYP11A1 ( gray ), CYP11A1 variant 1 ( blue ), and CYP11A1 variant 2 ( red ) expression in human peripheral steroidogenic tissues ( A ), human CNS tissues ( B ), and cell lines ( C ). Gene expression is shown as relative expression to β-actin. Data are presented as mean ± SD. CNS, central nervous system; qRT-PCR, quantitative RT-PCR.

    Journal: The Journal of Biological Chemistry

    Article Title: The neurosteroid pregnenolone is synthesized by a mitochondrial P450 enzyme other than CYP11A1 in human glial cells

    doi: 10.1016/j.jbc.2022.102110

    Figure Lengend Snippet: RNA Expression of CYP11A1 variants in the human CNS and peripheral tissues and cells . qRT-PCR analyses of total CYP11A1 ( gray ), CYP11A1 variant 1 ( blue ), and CYP11A1 variant 2 ( red ) expression in human peripheral steroidogenic tissues ( A ), human CNS tissues ( B ), and cell lines ( C ). Gene expression is shown as relative expression to β-actin. Data are presented as mean ± SD. CNS, central nervous system; qRT-PCR, quantitative RT-PCR.

    Article Snippet: The following primary antibodies were used: anti-CYP11A1 rabbit mAb (Cell Signaling Technology; #14217, 1:1000 dilution, antigen: human CYP11A1 N terminus), anti-CYP11A1 rabbit pAb (Proteintech; #13363-1-AP, 1:1000 dilution, antigen: CYP11A1 aa 1–300), anti-CYP11A1 rabbit pAb (Abcam; #ab75497, 1:500 dilution, antigen: human CYP11A1 aa 288–337), anti-CYP11A1 rabbit pAb (Abcam; #ab232763, 1:1000 dilution, antigen: CYP11A1 aa 350–521), anti-GAPDH rabbit mAb (Cell Signaling Technology; #2118, 1:2000 dilution), anti-FDXR rabbit pAb (Proteintech; #15584-1-AP, 1:1000 dilution), anti-ADX rabbit mAb (Abcam; #ab108257, 1:1000 dilution), antibeta actin mouse mAb (Abcam; #ab8226, 1:3000 dilution), anti-CYP reductase rabbit pAb (Abcam; #ab13513, 1:1000 dilution), and anti-Myc-tag rabbit mAb (Cell Signaling Technology; #2278S, 1:1000 dilution).

    Techniques: RNA Expression, Quantitative RT-PCR, Variant Assay, Expressing

    Localization of CYP11A1 isoforms. Representative confocal images of MGM-1 cells transfected with CYP11A1a-myc-DDK or CYP11A1b-myc-DDK expression vector. Cells transfected with empty plasmid were used as a negative control. Cells were stained with anti-myc-tag antibody ( green ), with mitochondria labeled with Mitotracker Red and nuclei stained with DAPI ( blue ). Images were taken at 63× magnification. CYP11A1a is strictly localized to the mitochondria. CYP11A1b appears more dispersed throughout the cell and has limited colocalization with mitochondria. DAPI, 4′,6-diamidino-2-phenylindole.

    Journal: The Journal of Biological Chemistry

    Article Title: The neurosteroid pregnenolone is synthesized by a mitochondrial P450 enzyme other than CYP11A1 in human glial cells

    doi: 10.1016/j.jbc.2022.102110

    Figure Lengend Snippet: Localization of CYP11A1 isoforms. Representative confocal images of MGM-1 cells transfected with CYP11A1a-myc-DDK or CYP11A1b-myc-DDK expression vector. Cells transfected with empty plasmid were used as a negative control. Cells were stained with anti-myc-tag antibody ( green ), with mitochondria labeled with Mitotracker Red and nuclei stained with DAPI ( blue ). Images were taken at 63× magnification. CYP11A1a is strictly localized to the mitochondria. CYP11A1b appears more dispersed throughout the cell and has limited colocalization with mitochondria. DAPI, 4′,6-diamidino-2-phenylindole.

    Article Snippet: The following primary antibodies were used: anti-CYP11A1 rabbit mAb (Cell Signaling Technology; #14217, 1:1000 dilution, antigen: human CYP11A1 N terminus), anti-CYP11A1 rabbit pAb (Proteintech; #13363-1-AP, 1:1000 dilution, antigen: CYP11A1 aa 1–300), anti-CYP11A1 rabbit pAb (Abcam; #ab75497, 1:500 dilution, antigen: human CYP11A1 aa 288–337), anti-CYP11A1 rabbit pAb (Abcam; #ab232763, 1:1000 dilution, antigen: CYP11A1 aa 350–521), anti-GAPDH rabbit mAb (Cell Signaling Technology; #2118, 1:2000 dilution), anti-FDXR rabbit pAb (Proteintech; #15584-1-AP, 1:1000 dilution), anti-ADX rabbit mAb (Abcam; #ab108257, 1:1000 dilution), antibeta actin mouse mAb (Abcam; #ab8226, 1:3000 dilution), anti-CYP reductase rabbit pAb (Abcam; #ab13513, 1:1000 dilution), and anti-Myc-tag rabbit mAb (Cell Signaling Technology; #2278S, 1:1000 dilution).

    Techniques: Transfection, Expressing, Plasmid Preparation, Negative Control, Staining, Labeling

    CYP11A1a but not CYP11A1b overexpression leads to increased pregnenolone production that can be inhibited by AMG . A and B , qRT-PCR analysis of CYP11A1 expression in MGM-1 cells transfected with empty plasmid (negative control) or expression vectors for CYP11A1a or CYP11A1b with Myc-DDK tag. Gene expression is shown as fold change versus negative control, normalized to the expression of β-actin. Data are presented as mean ± SD, N = 3. Each data point represents total RNA extracted from pooled cells from different passages. A , transfected MGM-1 cells showed more than 200-fold increase in total CYP11A1 mRNA expression. B , expression changes of CYP11A1 variant 1 exon 1 indicate that the correct CYP11A1 isoform had been expressed. C , representative immunoblots for myc-tag in transfected MGM-1 cells. β-Actin was used as a loading control. Thirty micrograms of protein were loaded into each lane. A band corresponding to CYP11A1a was observed at around 50 kDa and a band corresponding to CYP11A1b was observed at around 42 kDa. No specific bands could be observed in WT or transfection control MGM-1 cells. D and E , ELISA measurements of pregnenolone secreted by transfected MGM-1 cells, with or without 22(R)-hydroxycholesterol stimulation. F – I , ELISA measurements of pregnenolone secreted by transfected MGM-1 cells treated with different doses of AMG for 2 h under basal ( F and H ) and 22(R)-hydroxycholesterol stimulated ( G and I ) conditions. Each data point represents the average of one experiment, where each treatment was performed in triplicate within each experiment. Data are presented as mean ± SD, N = 3. Statistics performed compared to control ( D and E ) or to 0 mM AMG group ( F – I ). CYP11A1a+ cells synthesized significantly more pregnenolone than controls and CYP11A1b+ cells, which were significantly inhibited by AMG. CYP11A1b+ cells behaved similarly to WT MGM-1 cells for pregnenolone synthesis. (∗∗ p < 0.01, ∗∗∗ p < 0.001). AMG, aminoglutethimide; qRT-PCR, quantitative RT-PCR.

    Journal: The Journal of Biological Chemistry

    Article Title: The neurosteroid pregnenolone is synthesized by a mitochondrial P450 enzyme other than CYP11A1 in human glial cells

    doi: 10.1016/j.jbc.2022.102110

    Figure Lengend Snippet: CYP11A1a but not CYP11A1b overexpression leads to increased pregnenolone production that can be inhibited by AMG . A and B , qRT-PCR analysis of CYP11A1 expression in MGM-1 cells transfected with empty plasmid (negative control) or expression vectors for CYP11A1a or CYP11A1b with Myc-DDK tag. Gene expression is shown as fold change versus negative control, normalized to the expression of β-actin. Data are presented as mean ± SD, N = 3. Each data point represents total RNA extracted from pooled cells from different passages. A , transfected MGM-1 cells showed more than 200-fold increase in total CYP11A1 mRNA expression. B , expression changes of CYP11A1 variant 1 exon 1 indicate that the correct CYP11A1 isoform had been expressed. C , representative immunoblots for myc-tag in transfected MGM-1 cells. β-Actin was used as a loading control. Thirty micrograms of protein were loaded into each lane. A band corresponding to CYP11A1a was observed at around 50 kDa and a band corresponding to CYP11A1b was observed at around 42 kDa. No specific bands could be observed in WT or transfection control MGM-1 cells. D and E , ELISA measurements of pregnenolone secreted by transfected MGM-1 cells, with or without 22(R)-hydroxycholesterol stimulation. F – I , ELISA measurements of pregnenolone secreted by transfected MGM-1 cells treated with different doses of AMG for 2 h under basal ( F and H ) and 22(R)-hydroxycholesterol stimulated ( G and I ) conditions. Each data point represents the average of one experiment, where each treatment was performed in triplicate within each experiment. Data are presented as mean ± SD, N = 3. Statistics performed compared to control ( D and E ) or to 0 mM AMG group ( F – I ). CYP11A1a+ cells synthesized significantly more pregnenolone than controls and CYP11A1b+ cells, which were significantly inhibited by AMG. CYP11A1b+ cells behaved similarly to WT MGM-1 cells for pregnenolone synthesis. (∗∗ p < 0.01, ∗∗∗ p < 0.001). AMG, aminoglutethimide; qRT-PCR, quantitative RT-PCR.

    Article Snippet: The following primary antibodies were used: anti-CYP11A1 rabbit mAb (Cell Signaling Technology; #14217, 1:1000 dilution, antigen: human CYP11A1 N terminus), anti-CYP11A1 rabbit pAb (Proteintech; #13363-1-AP, 1:1000 dilution, antigen: CYP11A1 aa 1–300), anti-CYP11A1 rabbit pAb (Abcam; #ab75497, 1:500 dilution, antigen: human CYP11A1 aa 288–337), anti-CYP11A1 rabbit pAb (Abcam; #ab232763, 1:1000 dilution, antigen: CYP11A1 aa 350–521), anti-GAPDH rabbit mAb (Cell Signaling Technology; #2118, 1:2000 dilution), anti-FDXR rabbit pAb (Proteintech; #15584-1-AP, 1:1000 dilution), anti-ADX rabbit mAb (Abcam; #ab108257, 1:1000 dilution), antibeta actin mouse mAb (Abcam; #ab8226, 1:3000 dilution), anti-CYP reductase rabbit pAb (Abcam; #ab13513, 1:1000 dilution), and anti-Myc-tag rabbit mAb (Cell Signaling Technology; #2278S, 1:1000 dilution).

    Techniques: Over Expression, Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Negative Control, Variant Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Synthesized

    Effect of iron chelation on pregnenolone secretion in MGM-1 cells . ELISA measurements of secreted pregnenolone when MGM-1 WT ( A , B , E , and F ) and CYP11A1a+ ( C and D ) cells were pretreated with different doses of deferoxamine ( A – D ) or deferiprone ( E – F ) for 24 h. Each data point represents the average of one experiment, where each treatment was performed in triplicate within each experiment. Data are presented as mean ± SD, N = 3. Statistics performed compared to the no treatment group within each panel. Deferoxamine significantly inhibited pregnenolone production in CYP11A1+ cells and 22(R)-HC stimulated WT cells but had no statistically significant effect on WT cells under basal conditions. Deferiprone significantly inhibited pregnenolone production at high doses in basal and 22(R)-HC stimulated WT cells. (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). 22(R)-HC, 22(R)-hydroxycholesterol.

    Journal: The Journal of Biological Chemistry

    Article Title: The neurosteroid pregnenolone is synthesized by a mitochondrial P450 enzyme other than CYP11A1 in human glial cells

    doi: 10.1016/j.jbc.2022.102110

    Figure Lengend Snippet: Effect of iron chelation on pregnenolone secretion in MGM-1 cells . ELISA measurements of secreted pregnenolone when MGM-1 WT ( A , B , E , and F ) and CYP11A1a+ ( C and D ) cells were pretreated with different doses of deferoxamine ( A – D ) or deferiprone ( E – F ) for 24 h. Each data point represents the average of one experiment, where each treatment was performed in triplicate within each experiment. Data are presented as mean ± SD, N = 3. Statistics performed compared to the no treatment group within each panel. Deferoxamine significantly inhibited pregnenolone production in CYP11A1+ cells and 22(R)-HC stimulated WT cells but had no statistically significant effect on WT cells under basal conditions. Deferiprone significantly inhibited pregnenolone production at high doses in basal and 22(R)-HC stimulated WT cells. (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). 22(R)-HC, 22(R)-hydroxycholesterol.

    Article Snippet: The following primary antibodies were used: anti-CYP11A1 rabbit mAb (Cell Signaling Technology; #14217, 1:1000 dilution, antigen: human CYP11A1 N terminus), anti-CYP11A1 rabbit pAb (Proteintech; #13363-1-AP, 1:1000 dilution, antigen: CYP11A1 aa 1–300), anti-CYP11A1 rabbit pAb (Abcam; #ab75497, 1:500 dilution, antigen: human CYP11A1 aa 288–337), anti-CYP11A1 rabbit pAb (Abcam; #ab232763, 1:1000 dilution, antigen: CYP11A1 aa 350–521), anti-GAPDH rabbit mAb (Cell Signaling Technology; #2118, 1:2000 dilution), anti-FDXR rabbit pAb (Proteintech; #15584-1-AP, 1:1000 dilution), anti-ADX rabbit mAb (Abcam; #ab108257, 1:1000 dilution), antibeta actin mouse mAb (Abcam; #ab8226, 1:3000 dilution), anti-CYP reductase rabbit pAb (Abcam; #ab13513, 1:1000 dilution), and anti-Myc-tag rabbit mAb (Cell Signaling Technology; #2278S, 1:1000 dilution).

    Techniques: Enzyme-linked Immunosorbent Assay

    PS-MPs treatment downregulated the expression of steroidogenic enzymes and StAR in testes. Mice were given drinking water comprising PS-MPs of various sizes for 180 sustained days. A , B The expression of 3β-HSD, 17β-HSD, P450scc, P450c17, and StAR in testes was measured by western blotting. The expression levels were quantified with ImageJ (n = 3). Data are expressed as means ± SD. * P < 0.05, ** P < 0.01 vs. control. # P < 0.05, ## P < 0.01 vs. 100 μg/L group. C , D The expression of StAR in mouse testicular tissues of the control group and 1000 μg/L group was tested by immunofluorescence staining. Testicular tissues were stained with StAR (red) and DAPI (blue), arrows: positive expression (scale bar = 50 µm). The expression levels of StAR were quantified with ImageJ (n = 3). Data are expressed as means ± SD. ** P < 0.01 vs. control. # P < 0.05, *** P < 0.001 vs. control

    Journal: Particle and Fibre Toxicology

    Article Title: Chronic exposure to polystyrene microplastics induced male reproductive toxicity and decreased testosterone levels via the LH-mediated LHR/cAMP/PKA/StAR pathway

    doi: 10.1186/s12989-022-00453-2

    Figure Lengend Snippet: PS-MPs treatment downregulated the expression of steroidogenic enzymes and StAR in testes. Mice were given drinking water comprising PS-MPs of various sizes for 180 sustained days. A , B The expression of 3β-HSD, 17β-HSD, P450scc, P450c17, and StAR in testes was measured by western blotting. The expression levels were quantified with ImageJ (n = 3). Data are expressed as means ± SD. * P < 0.05, ** P < 0.01 vs. control. # P < 0.05, ## P < 0.01 vs. 100 μg/L group. C , D The expression of StAR in mouse testicular tissues of the control group and 1000 μg/L group was tested by immunofluorescence staining. Testicular tissues were stained with StAR (red) and DAPI (blue), arrows: positive expression (scale bar = 50 µm). The expression levels of StAR were quantified with ImageJ (n = 3). Data are expressed as means ± SD. ** P < 0.01 vs. control. # P < 0.05, *** P < 0.001 vs. control

    Article Snippet: Transferred blots were incubated with rabbit anti-CYP11A (P450scc) (Proteintech Group, Rosemont, IL, USA), rabbit anti-CYP17A (P450c17) (Proteintech Group, Rosemont, IL, USA), rabbit anti-StAR (Cell Signaling Technology, USA), mouse anti-HSD3β (Proteintech Group, Rosemont, IL, USA), rabbit anti-HSD17β2 (Proteintech Group, Rosemont, IL, USA), rabbit anti-LHR (Proteintech Group, Rosemont, IL, USA), and mouse anti-GAPDH (Proteintech Group, Rosemont, IL, USA) overnight at 4° C. Specific information on the antibodies was shown in Table .

    Techniques: Expressing, Western Blot, Immunofluorescence, Staining

    The expression of steroidogenic enzymes and StAR decreased in Leydig cells after PS-MPs treatment. Primary Leydig cells were exposed to 0.5 μm PS-MPs for 24 h at various concentrations as indicated. A The content of testosterone in supernatant was determined by ELISA assays (n = 3). B The mRNA expression levels of 3β-HSD, 17β-HSD, P450scc, P450c17, and StAR in Leydig cells after treatment with PS-MPs were determined by qRT-PCR (n = 3). C , D The expression of 3β-HSD, 17β-HSD, P450scc, and P450c17 in cells was measured by western blotting. The expression levels were quantified with ImageJ (n = 3). Data are expressed as means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. control

    Journal: Particle and Fibre Toxicology

    Article Title: Chronic exposure to polystyrene microplastics induced male reproductive toxicity and decreased testosterone levels via the LH-mediated LHR/cAMP/PKA/StAR pathway

    doi: 10.1186/s12989-022-00453-2

    Figure Lengend Snippet: The expression of steroidogenic enzymes and StAR decreased in Leydig cells after PS-MPs treatment. Primary Leydig cells were exposed to 0.5 μm PS-MPs for 24 h at various concentrations as indicated. A The content of testosterone in supernatant was determined by ELISA assays (n = 3). B The mRNA expression levels of 3β-HSD, 17β-HSD, P450scc, P450c17, and StAR in Leydig cells after treatment with PS-MPs were determined by qRT-PCR (n = 3). C , D The expression of 3β-HSD, 17β-HSD, P450scc, and P450c17 in cells was measured by western blotting. The expression levels were quantified with ImageJ (n = 3). Data are expressed as means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. control

    Article Snippet: Transferred blots were incubated with rabbit anti-CYP11A (P450scc) (Proteintech Group, Rosemont, IL, USA), rabbit anti-CYP17A (P450c17) (Proteintech Group, Rosemont, IL, USA), rabbit anti-StAR (Cell Signaling Technology, USA), mouse anti-HSD3β (Proteintech Group, Rosemont, IL, USA), rabbit anti-HSD17β2 (Proteintech Group, Rosemont, IL, USA), rabbit anti-LHR (Proteintech Group, Rosemont, IL, USA), and mouse anti-GAPDH (Proteintech Group, Rosemont, IL, USA) overnight at 4° C. Specific information on the antibodies was shown in Table .

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

    Overexpression of LHR alleviated the reduction in StAR, steroidogenic enzymes and testosterone levels induced by PS-MPs treatment in primary Leydig cells. A Primary cells were infected with LHR or empty vector with lentivirus for 72 h. The mRNA levels of LHR were tested by qRT-PCR (n = 3; *** P < 0.001 vs. LV-NC group). B Primary cells were infected with LHR or empty vector with lentivirus for 72 h. Then, the cells were exposed to 0.5 μm PS-MPs at a concentration of 0.2 mg/mL for 24 h. The testosterone content in the supernatant was examined by ELISA (n = 3; *** P < 0.001 vs. LV-NC group; ## P < 0.01 vs. LV-NC + PS-MPs group). C , D Primary cells were infected with LHR or empty vector with lentivirus for 72 h. Then, the cells were treated with 0.5 μm PS-MPs with a concentration at 0.2 mg/mL for 24 h. The expression of 3β-HSD, 17β-HSD, P450scc, P450c17, StAR, and LHR in cells was measured by western blotting. The expression levels were quantified with ImageJ (n = 3; * P < 0.05, ** P < 0.01 vs. LV-NC group; # P < 0.05, ## P < 0.01, ### P < 0.01 vs. LV-NC + PS-MPs group)

    Journal: Particle and Fibre Toxicology

    Article Title: Chronic exposure to polystyrene microplastics induced male reproductive toxicity and decreased testosterone levels via the LH-mediated LHR/cAMP/PKA/StAR pathway

    doi: 10.1186/s12989-022-00453-2

    Figure Lengend Snippet: Overexpression of LHR alleviated the reduction in StAR, steroidogenic enzymes and testosterone levels induced by PS-MPs treatment in primary Leydig cells. A Primary cells were infected with LHR or empty vector with lentivirus for 72 h. The mRNA levels of LHR were tested by qRT-PCR (n = 3; *** P < 0.001 vs. LV-NC group). B Primary cells were infected with LHR or empty vector with lentivirus for 72 h. Then, the cells were exposed to 0.5 μm PS-MPs at a concentration of 0.2 mg/mL for 24 h. The testosterone content in the supernatant was examined by ELISA (n = 3; *** P < 0.001 vs. LV-NC group; ## P < 0.01 vs. LV-NC + PS-MPs group). C , D Primary cells were infected with LHR or empty vector with lentivirus for 72 h. Then, the cells were treated with 0.5 μm PS-MPs with a concentration at 0.2 mg/mL for 24 h. The expression of 3β-HSD, 17β-HSD, P450scc, P450c17, StAR, and LHR in cells was measured by western blotting. The expression levels were quantified with ImageJ (n = 3; * P < 0.05, ** P < 0.01 vs. LV-NC group; # P < 0.05, ## P < 0.01, ### P < 0.01 vs. LV-NC + PS-MPs group)

    Article Snippet: Transferred blots were incubated with rabbit anti-CYP11A (P450scc) (Proteintech Group, Rosemont, IL, USA), rabbit anti-CYP17A (P450c17) (Proteintech Group, Rosemont, IL, USA), rabbit anti-StAR (Cell Signaling Technology, USA), mouse anti-HSD3β (Proteintech Group, Rosemont, IL, USA), rabbit anti-HSD17β2 (Proteintech Group, Rosemont, IL, USA), rabbit anti-LHR (Proteintech Group, Rosemont, IL, USA), and mouse anti-GAPDH (Proteintech Group, Rosemont, IL, USA) overnight at 4° C. Specific information on the antibodies was shown in Table .

    Techniques: Over Expression, Infection, Plasmid Preparation, Quantitative RT-PCR, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

    Specifications of primary antibodies

    Journal: Particle and Fibre Toxicology

    Article Title: Chronic exposure to polystyrene microplastics induced male reproductive toxicity and decreased testosterone levels via the LH-mediated LHR/cAMP/PKA/StAR pathway

    doi: 10.1186/s12989-022-00453-2

    Figure Lengend Snippet: Specifications of primary antibodies

    Article Snippet: Transferred blots were incubated with rabbit anti-CYP11A (P450scc) (Proteintech Group, Rosemont, IL, USA), rabbit anti-CYP17A (P450c17) (Proteintech Group, Rosemont, IL, USA), rabbit anti-StAR (Cell Signaling Technology, USA), mouse anti-HSD3β (Proteintech Group, Rosemont, IL, USA), rabbit anti-HSD17β2 (Proteintech Group, Rosemont, IL, USA), rabbit anti-LHR (Proteintech Group, Rosemont, IL, USA), and mouse anti-GAPDH (Proteintech Group, Rosemont, IL, USA) overnight at 4° C. Specific information on the antibodies was shown in Table .

    Techniques: