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aqp3 protein  (Boster Bio)


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    Boster Bio aqp3 protein
    Effect of AHE on <t>AQP3</t> expression. ( a ) HaCaT cells were treated with different concentrations of AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were quantitated by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) AQP3 promoter assay. pAQP3-Luc(−1090/+16) reporter was transfected into HaCaT cells. After 48 h, cells were treated with different concentrations of AHE. After 8–12 h, cells were collected and luciferase activities were measured. ( d ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( e ) HaCaT cells were treated as in ( d ). AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ( f ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. <t>AQP3</t> <t>protein</t> levels were analyzed by immunoblotting. GAPDH expression was used as an internal control. ( g ) Flow cytometry. HaCaT cells were treated with AHE (10 and 20 μg/mL) and the percentage of AQP3 positive cells were measured by flow cytometry. Full-length gels ( a and d ) and blots ( f ) are presented in Supplemental Fig. .
    Aqp3 Protein, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aqp3 protein/product/Boster Bio
    Average 90 stars, based on 1 article reviews
    aqp3 protein - by Bioz Stars, 2026-02
    90/100 stars

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    1) Product Images from "Agerarin, identified from Ageratum houstonianum , stimulates circadian CLOCK-mediated aquaporin-3 gene expression in HaCaT keratinocytes"

    Article Title: Agerarin, identified from Ageratum houstonianum , stimulates circadian CLOCK-mediated aquaporin-3 gene expression in HaCaT keratinocytes

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-11642-x

    Effect of AHE on AQP3 expression. ( a ) HaCaT cells were treated with different concentrations of AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were quantitated by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) AQP3 promoter assay. pAQP3-Luc(−1090/+16) reporter was transfected into HaCaT cells. After 48 h, cells were treated with different concentrations of AHE. After 8–12 h, cells were collected and luciferase activities were measured. ( d ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( e ) HaCaT cells were treated as in ( d ). AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ( f ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 protein levels were analyzed by immunoblotting. GAPDH expression was used as an internal control. ( g ) Flow cytometry. HaCaT cells were treated with AHE (10 and 20 μg/mL) and the percentage of AQP3 positive cells were measured by flow cytometry. Full-length gels ( a and d ) and blots ( f ) are presented in Supplemental Fig. .
    Figure Legend Snippet: Effect of AHE on AQP3 expression. ( a ) HaCaT cells were treated with different concentrations of AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were quantitated by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) AQP3 promoter assay. pAQP3-Luc(−1090/+16) reporter was transfected into HaCaT cells. After 48 h, cells were treated with different concentrations of AHE. After 8–12 h, cells were collected and luciferase activities were measured. ( d ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( e ) HaCaT cells were treated as in ( d ). AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ( f ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 protein levels were analyzed by immunoblotting. GAPDH expression was used as an internal control. ( g ) Flow cytometry. HaCaT cells were treated with AHE (10 and 20 μg/mL) and the percentage of AQP3 positive cells were measured by flow cytometry. Full-length gels ( a and d ) and blots ( f ) are presented in Supplemental Fig. .

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR, Promoter Assay, Transfection, Luciferase, Western Blot, Flow Cytometry

    Role of MAPK signaling in AHE-induced AQP3 expression. ( a ) HaCaT cells were treated with 20 μg/mL AHE for various lengths of time. Phosphorylation status of MAPKs were analyzed by immunoblotting. GAPDH antibody was used as an internal control to show equal protein loading. ( b ) HaCaT cells were either untreated or pretreated with U0126 (10 μM), SB203580 (20 μM), or SP600125 (25 μM) before addition of 20 μg/mL AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( c ) HaCaT cells were treated as in ( b ). AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ns, not significant. Full-length blots ( a ) and gels ( b ) are presented in Supplemental Fig. .
    Figure Legend Snippet: Role of MAPK signaling in AHE-induced AQP3 expression. ( a ) HaCaT cells were treated with 20 μg/mL AHE for various lengths of time. Phosphorylation status of MAPKs were analyzed by immunoblotting. GAPDH antibody was used as an internal control to show equal protein loading. ( b ) HaCaT cells were either untreated or pretreated with U0126 (10 μM), SB203580 (20 μM), or SP600125 (25 μM) before addition of 20 μg/mL AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( c ) HaCaT cells were treated as in ( b ). AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ns, not significant. Full-length blots ( a ) and gels ( b ) are presented in Supplemental Fig. .

    Techniques Used: Expressing, Phospho-proteomics, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    Effect of UVB on the suppression of AQP3 expression. ( a ) RT-PCR. HaCaT cells were irradiated with different doses of UVB for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were irradiated with UVB as in ( c ). AQP3 mRNA levels were quantitated by qRT-PCR. The mRNA expression were normalized to the GAPDH mRNA. Full-length gels ( a and c ) are presented in Supplemental Fig. .
    Figure Legend Snippet: Effect of UVB on the suppression of AQP3 expression. ( a ) RT-PCR. HaCaT cells were irradiated with different doses of UVB for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were irradiated with UVB as in ( c ). AQP3 mRNA levels were quantitated by qRT-PCR. The mRNA expression were normalized to the GAPDH mRNA. Full-length gels ( a and c ) are presented in Supplemental Fig. .

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Irradiation, Control, Quantitative RT-PCR

    Effect of AHE on UVB-induced suppression of AQP3 expression. ( a ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of different concentrations of AHE. After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and qRT-PCR was performed. GAPDH mRNA level was used for normalization. ( c ) HaCaT cells were transfected with 0.2 μg of promoter reporter pAQP3-Luc(−1090/+16), followed by irradiation with UVB (30 mJ/cm 2 ) in the absence or presence of 20 μg/mL AHE at 24 h post-transfection. After 8–12 h, luciferase activities were measured. ( d ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of AHE (20 and 10 μg/mL). After 24 h, AQP3 protein level was detected by immunoblotting. GAPDH was used as an internal control. ( e ) HaCaT cells cultured on a coverglass were treated as in ( d ). AQP3 antibody was incubated for 2 h, followed by AlexaFluor 555-conjugated (red signal) secondary antibody for additional 30 min. Nuclear DNA was stained with 0.1 μg/mL Hoechst 33258 (blue signal) for 10 min. Fluorescence-positive cells were examined under an EVOSf1 ® fluorescence microscope. Full-length gels ( a ) and blots ( d ) are presented in Supplemental Fig. .
    Figure Legend Snippet: Effect of AHE on UVB-induced suppression of AQP3 expression. ( a ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of different concentrations of AHE. After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and qRT-PCR was performed. GAPDH mRNA level was used for normalization. ( c ) HaCaT cells were transfected with 0.2 μg of promoter reporter pAQP3-Luc(−1090/+16), followed by irradiation with UVB (30 mJ/cm 2 ) in the absence or presence of 20 μg/mL AHE at 24 h post-transfection. After 8–12 h, luciferase activities were measured. ( d ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of AHE (20 and 10 μg/mL). After 24 h, AQP3 protein level was detected by immunoblotting. GAPDH was used as an internal control. ( e ) HaCaT cells cultured on a coverglass were treated as in ( d ). AQP3 antibody was incubated for 2 h, followed by AlexaFluor 555-conjugated (red signal) secondary antibody for additional 30 min. Nuclear DNA was stained with 0.1 μg/mL Hoechst 33258 (blue signal) for 10 min. Fluorescence-positive cells were examined under an EVOSf1 ® fluorescence microscope. Full-length gels ( a ) and blots ( d ) are presented in Supplemental Fig. .

    Techniques Used: Expressing, Irradiation, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR, Transfection, Luciferase, Western Blot, Cell Culture, Incubation, Staining, Fluorescence, Microscopy

    Effect of agerarin on the expression of AQP3 . ( a ) HaCaT cells were treated with different concentrations of AHE or agerarin. After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ** p < 0.0001 compared to vehicle control (0 μg/mL). ( c ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of AHE or agerarin (each 20 μg/mL). After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( c ) and qRT-PCR was performed. GAPDH mRNA level was used for normalization. Full-length gels ( a and c ) are presented in Supplemental Fig. .
    Figure Legend Snippet: Effect of agerarin on the expression of AQP3 . ( a ) HaCaT cells were treated with different concentrations of AHE or agerarin. After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ** p < 0.0001 compared to vehicle control (0 μg/mL). ( c ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of AHE or agerarin (each 20 μg/mL). After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( c ) and qRT-PCR was performed. GAPDH mRNA level was used for normalization. Full-length gels ( a and c ) are presented in Supplemental Fig. .

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR, Irradiation

    Effect of agerarin on the expression of CLOCK . ( a ) HaCaT cells were transfected with 0.2 μg of promoter construct pAQP3-Luc(−1090/+16) along with different concentrations of expression plasmids for CLOCK , pcDNA3.1/Clock. After 24 h, cells were collected and assayed for luciferase activity. ( b ) HaCaT cells were transfected with 0.2 µg of a full-length (−1090/+16) or 5′-deletion construct (−198/+16) along with 0.1 μg of empty vector (pcDNA3,1) or pcDNA3.1/Clock plasmids, and measured luciferase activity after 24 h. ( c ) HaCaT cells were treated with agerarin (20 μg/mL) for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( b ) and AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( e ) HaCaT cells were treated with 20 μg/mL agerarin for various time periods. AQP3 protein level was detected by immunoblotting. GAPDH was used as an internal control. ( f ) CLOCK promoter assay. HaCaT cells were transfected with 0.2 μg of promoter reporter pClock-Luc(−1000/−1). After 48 h, HaCaT cells were treated with different concentrations of agerarin. After 12 h, luciferase activities were measured. Full-length gels ( c ) and blots ( e ) are presented in Supplemental Fig. .
    Figure Legend Snippet: Effect of agerarin on the expression of CLOCK . ( a ) HaCaT cells were transfected with 0.2 μg of promoter construct pAQP3-Luc(−1090/+16) along with different concentrations of expression plasmids for CLOCK , pcDNA3.1/Clock. After 24 h, cells were collected and assayed for luciferase activity. ( b ) HaCaT cells were transfected with 0.2 µg of a full-length (−1090/+16) or 5′-deletion construct (−198/+16) along with 0.1 μg of empty vector (pcDNA3,1) or pcDNA3.1/Clock plasmids, and measured luciferase activity after 24 h. ( c ) HaCaT cells were treated with agerarin (20 μg/mL) for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( b ) and AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( e ) HaCaT cells were treated with 20 μg/mL agerarin for various time periods. AQP3 protein level was detected by immunoblotting. GAPDH was used as an internal control. ( f ) CLOCK promoter assay. HaCaT cells were transfected with 0.2 μg of promoter reporter pClock-Luc(−1000/−1). After 48 h, HaCaT cells were treated with different concentrations of agerarin. After 12 h, luciferase activities were measured. Full-length gels ( c ) and blots ( e ) are presented in Supplemental Fig. .

    Techniques Used: Expressing, Transfection, Construct, Luciferase, Activity Assay, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR, Western Blot, Promoter Assay

    Effect of CLOCK silencing on agerarin-induced AQP3 expression. ( a ) HaCaT/shCont and HaCaT/shClock cells were treated with agerarin (each 20 μg/mL) for 12 h. CLOCK mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and CLOCK mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) HaCaT/shCont and HaCaT/shClock cells were treated with agerarin (20 μg/mL). After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( c ) and AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. Full-length gels ( a and c ) are presented in Supplemental Fig. .
    Figure Legend Snippet: Effect of CLOCK silencing on agerarin-induced AQP3 expression. ( a ) HaCaT/shCont and HaCaT/shClock cells were treated with agerarin (each 20 μg/mL) for 12 h. CLOCK mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and CLOCK mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) HaCaT/shCont and HaCaT/shClock cells were treated with agerarin (20 μg/mL). After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( c ) and AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. Full-length gels ( a and c ) are presented in Supplemental Fig. .

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR



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    aqp3 protein  (Boster Bio)
    90
    Boster Bio aqp3 protein
    Effect of AHE on <t>AQP3</t> expression. ( a ) HaCaT cells were treated with different concentrations of AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were quantitated by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) AQP3 promoter assay. pAQP3-Luc(−1090/+16) reporter was transfected into HaCaT cells. After 48 h, cells were treated with different concentrations of AHE. After 8–12 h, cells were collected and luciferase activities were measured. ( d ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( e ) HaCaT cells were treated as in ( d ). AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ( f ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. <t>AQP3</t> <t>protein</t> levels were analyzed by immunoblotting. GAPDH expression was used as an internal control. ( g ) Flow cytometry. HaCaT cells were treated with AHE (10 and 20 μg/mL) and the percentage of AQP3 positive cells were measured by flow cytometry. Full-length gels ( a and d ) and blots ( f ) are presented in Supplemental Fig. .
    Aqp3 Protein, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aqp3 protein/product/Boster Bio
    Average 90 stars, based on 1 article reviews
    aqp3 protein - by Bioz Stars, 2026-02
    90/100 stars
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    Effect of AHE on AQP3 expression. ( a ) HaCaT cells were treated with different concentrations of AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were quantitated by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) AQP3 promoter assay. pAQP3-Luc(−1090/+16) reporter was transfected into HaCaT cells. After 48 h, cells were treated with different concentrations of AHE. After 8–12 h, cells were collected and luciferase activities were measured. ( d ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( e ) HaCaT cells were treated as in ( d ). AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ( f ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 protein levels were analyzed by immunoblotting. GAPDH expression was used as an internal control. ( g ) Flow cytometry. HaCaT cells were treated with AHE (10 and 20 μg/mL) and the percentage of AQP3 positive cells were measured by flow cytometry. Full-length gels ( a and d ) and blots ( f ) are presented in Supplemental Fig. .

    Journal: Scientific Reports

    Article Title: Agerarin, identified from Ageratum houstonianum , stimulates circadian CLOCK-mediated aquaporin-3 gene expression in HaCaT keratinocytes

    doi: 10.1038/s41598-017-11642-x

    Figure Lengend Snippet: Effect of AHE on AQP3 expression. ( a ) HaCaT cells were treated with different concentrations of AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were quantitated by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) AQP3 promoter assay. pAQP3-Luc(−1090/+16) reporter was transfected into HaCaT cells. After 48 h, cells were treated with different concentrations of AHE. After 8–12 h, cells were collected and luciferase activities were measured. ( d ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( e ) HaCaT cells were treated as in ( d ). AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ( f ) HaCaT cells were treated with 20 μg/mL AHE for various time periods. AQP3 protein levels were analyzed by immunoblotting. GAPDH expression was used as an internal control. ( g ) Flow cytometry. HaCaT cells were treated with AHE (10 and 20 μg/mL) and the percentage of AQP3 positive cells were measured by flow cytometry. Full-length gels ( a and d ) and blots ( f ) are presented in Supplemental Fig. .

    Article Snippet: AQP3 protein was detected using 1:500 diluted ant-AQP3 antibody (BOSTER).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR, Promoter Assay, Transfection, Luciferase, Western Blot, Flow Cytometry

    Role of MAPK signaling in AHE-induced AQP3 expression. ( a ) HaCaT cells were treated with 20 μg/mL AHE for various lengths of time. Phosphorylation status of MAPKs were analyzed by immunoblotting. GAPDH antibody was used as an internal control to show equal protein loading. ( b ) HaCaT cells were either untreated or pretreated with U0126 (10 μM), SB203580 (20 μM), or SP600125 (25 μM) before addition of 20 μg/mL AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( c ) HaCaT cells were treated as in ( b ). AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ns, not significant. Full-length blots ( a ) and gels ( b ) are presented in Supplemental Fig. .

    Journal: Scientific Reports

    Article Title: Agerarin, identified from Ageratum houstonianum , stimulates circadian CLOCK-mediated aquaporin-3 gene expression in HaCaT keratinocytes

    doi: 10.1038/s41598-017-11642-x

    Figure Lengend Snippet: Role of MAPK signaling in AHE-induced AQP3 expression. ( a ) HaCaT cells were treated with 20 μg/mL AHE for various lengths of time. Phosphorylation status of MAPKs were analyzed by immunoblotting. GAPDH antibody was used as an internal control to show equal protein loading. ( b ) HaCaT cells were either untreated or pretreated with U0126 (10 μM), SB203580 (20 μM), or SP600125 (25 μM) before addition of 20 μg/mL AHE for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( c ) HaCaT cells were treated as in ( b ). AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ns, not significant. Full-length blots ( a ) and gels ( b ) are presented in Supplemental Fig. .

    Article Snippet: AQP3 protein was detected using 1:500 diluted ant-AQP3 antibody (BOSTER).

    Techniques: Expressing, Phospho-proteomics, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    Effect of UVB on the suppression of AQP3 expression. ( a ) RT-PCR. HaCaT cells were irradiated with different doses of UVB for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were irradiated with UVB as in ( c ). AQP3 mRNA levels were quantitated by qRT-PCR. The mRNA expression were normalized to the GAPDH mRNA. Full-length gels ( a and c ) are presented in Supplemental Fig. .

    Journal: Scientific Reports

    Article Title: Agerarin, identified from Ageratum houstonianum , stimulates circadian CLOCK-mediated aquaporin-3 gene expression in HaCaT keratinocytes

    doi: 10.1038/s41598-017-11642-x

    Figure Lengend Snippet: Effect of UVB on the suppression of AQP3 expression. ( a ) RT-PCR. HaCaT cells were irradiated with different doses of UVB for 24 h. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated with as in ( a ). AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were irradiated with UVB as in ( c ). AQP3 mRNA levels were quantitated by qRT-PCR. The mRNA expression were normalized to the GAPDH mRNA. Full-length gels ( a and c ) are presented in Supplemental Fig. .

    Article Snippet: AQP3 protein was detected using 1:500 diluted ant-AQP3 antibody (BOSTER).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Irradiation, Control, Quantitative RT-PCR

    Effect of AHE on UVB-induced suppression of AQP3 expression. ( a ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of different concentrations of AHE. After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and qRT-PCR was performed. GAPDH mRNA level was used for normalization. ( c ) HaCaT cells were transfected with 0.2 μg of promoter reporter pAQP3-Luc(−1090/+16), followed by irradiation with UVB (30 mJ/cm 2 ) in the absence or presence of 20 μg/mL AHE at 24 h post-transfection. After 8–12 h, luciferase activities were measured. ( d ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of AHE (20 and 10 μg/mL). After 24 h, AQP3 protein level was detected by immunoblotting. GAPDH was used as an internal control. ( e ) HaCaT cells cultured on a coverglass were treated as in ( d ). AQP3 antibody was incubated for 2 h, followed by AlexaFluor 555-conjugated (red signal) secondary antibody for additional 30 min. Nuclear DNA was stained with 0.1 μg/mL Hoechst 33258 (blue signal) for 10 min. Fluorescence-positive cells were examined under an EVOSf1 ® fluorescence microscope. Full-length gels ( a ) and blots ( d ) are presented in Supplemental Fig. .

    Journal: Scientific Reports

    Article Title: Agerarin, identified from Ageratum houstonianum , stimulates circadian CLOCK-mediated aquaporin-3 gene expression in HaCaT keratinocytes

    doi: 10.1038/s41598-017-11642-x

    Figure Lengend Snippet: Effect of AHE on UVB-induced suppression of AQP3 expression. ( a ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of different concentrations of AHE. After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and qRT-PCR was performed. GAPDH mRNA level was used for normalization. ( c ) HaCaT cells were transfected with 0.2 μg of promoter reporter pAQP3-Luc(−1090/+16), followed by irradiation with UVB (30 mJ/cm 2 ) in the absence or presence of 20 μg/mL AHE at 24 h post-transfection. After 8–12 h, luciferase activities were measured. ( d ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of AHE (20 and 10 μg/mL). After 24 h, AQP3 protein level was detected by immunoblotting. GAPDH was used as an internal control. ( e ) HaCaT cells cultured on a coverglass were treated as in ( d ). AQP3 antibody was incubated for 2 h, followed by AlexaFluor 555-conjugated (red signal) secondary antibody for additional 30 min. Nuclear DNA was stained with 0.1 μg/mL Hoechst 33258 (blue signal) for 10 min. Fluorescence-positive cells were examined under an EVOSf1 ® fluorescence microscope. Full-length gels ( a ) and blots ( d ) are presented in Supplemental Fig. .

    Article Snippet: AQP3 protein was detected using 1:500 diluted ant-AQP3 antibody (BOSTER).

    Techniques: Expressing, Irradiation, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR, Transfection, Luciferase, Western Blot, Cell Culture, Incubation, Staining, Fluorescence, Microscopy

    Effect of agerarin on the expression of AQP3 . ( a ) HaCaT cells were treated with different concentrations of AHE or agerarin. After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ** p < 0.0001 compared to vehicle control (0 μg/mL). ( c ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of AHE or agerarin (each 20 μg/mL). After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( c ) and qRT-PCR was performed. GAPDH mRNA level was used for normalization. Full-length gels ( a and c ) are presented in Supplemental Fig. .

    Journal: Scientific Reports

    Article Title: Agerarin, identified from Ageratum houstonianum , stimulates circadian CLOCK-mediated aquaporin-3 gene expression in HaCaT keratinocytes

    doi: 10.1038/s41598-017-11642-x

    Figure Lengend Snippet: Effect of agerarin on the expression of AQP3 . ( a ) HaCaT cells were treated with different concentrations of AHE or agerarin. After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and AQP3 mRNA levels were measured by qRT-PCR. The relative fold changes were normalized to the expression of GAPDH mRNA. ** p < 0.0001 compared to vehicle control (0 μg/mL). ( c ) HaCaT cells were irradiated with UVB (30 mJ/cm 2 ) in the absence or presence of AHE or agerarin (each 20 μg/mL). After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( c ) and qRT-PCR was performed. GAPDH mRNA level was used for normalization. Full-length gels ( a and c ) are presented in Supplemental Fig. .

    Article Snippet: AQP3 protein was detected using 1:500 diluted ant-AQP3 antibody (BOSTER).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR, Irradiation

    Effect of agerarin on the expression of CLOCK . ( a ) HaCaT cells were transfected with 0.2 μg of promoter construct pAQP3-Luc(−1090/+16) along with different concentrations of expression plasmids for CLOCK , pcDNA3.1/Clock. After 24 h, cells were collected and assayed for luciferase activity. ( b ) HaCaT cells were transfected with 0.2 µg of a full-length (−1090/+16) or 5′-deletion construct (−198/+16) along with 0.1 μg of empty vector (pcDNA3,1) or pcDNA3.1/Clock plasmids, and measured luciferase activity after 24 h. ( c ) HaCaT cells were treated with agerarin (20 μg/mL) for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( b ) and AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( e ) HaCaT cells were treated with 20 μg/mL agerarin for various time periods. AQP3 protein level was detected by immunoblotting. GAPDH was used as an internal control. ( f ) CLOCK promoter assay. HaCaT cells were transfected with 0.2 μg of promoter reporter pClock-Luc(−1000/−1). After 48 h, HaCaT cells were treated with different concentrations of agerarin. After 12 h, luciferase activities were measured. Full-length gels ( c ) and blots ( e ) are presented in Supplemental Fig. .

    Journal: Scientific Reports

    Article Title: Agerarin, identified from Ageratum houstonianum , stimulates circadian CLOCK-mediated aquaporin-3 gene expression in HaCaT keratinocytes

    doi: 10.1038/s41598-017-11642-x

    Figure Lengend Snippet: Effect of agerarin on the expression of CLOCK . ( a ) HaCaT cells were transfected with 0.2 μg of promoter construct pAQP3-Luc(−1090/+16) along with different concentrations of expression plasmids for CLOCK , pcDNA3.1/Clock. After 24 h, cells were collected and assayed for luciferase activity. ( b ) HaCaT cells were transfected with 0.2 µg of a full-length (−1090/+16) or 5′-deletion construct (−198/+16) along with 0.1 μg of empty vector (pcDNA3,1) or pcDNA3.1/Clock plasmids, and measured luciferase activity after 24 h. ( c ) HaCaT cells were treated with agerarin (20 μg/mL) for various time periods. AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( b ) and AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( e ) HaCaT cells were treated with 20 μg/mL agerarin for various time periods. AQP3 protein level was detected by immunoblotting. GAPDH was used as an internal control. ( f ) CLOCK promoter assay. HaCaT cells were transfected with 0.2 μg of promoter reporter pClock-Luc(−1000/−1). After 48 h, HaCaT cells were treated with different concentrations of agerarin. After 12 h, luciferase activities were measured. Full-length gels ( c ) and blots ( e ) are presented in Supplemental Fig. .

    Article Snippet: AQP3 protein was detected using 1:500 diluted ant-AQP3 antibody (BOSTER).

    Techniques: Expressing, Transfection, Construct, Luciferase, Activity Assay, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR, Western Blot, Promoter Assay

    Effect of CLOCK silencing on agerarin-induced AQP3 expression. ( a ) HaCaT/shCont and HaCaT/shClock cells were treated with agerarin (each 20 μg/mL) for 12 h. CLOCK mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and CLOCK mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) HaCaT/shCont and HaCaT/shClock cells were treated with agerarin (20 μg/mL). After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( c ) and AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. Full-length gels ( a and c ) are presented in Supplemental Fig. .

    Journal: Scientific Reports

    Article Title: Agerarin, identified from Ageratum houstonianum , stimulates circadian CLOCK-mediated aquaporin-3 gene expression in HaCaT keratinocytes

    doi: 10.1038/s41598-017-11642-x

    Figure Lengend Snippet: Effect of CLOCK silencing on agerarin-induced AQP3 expression. ( a ) HaCaT/shCont and HaCaT/shClock cells were treated with agerarin (each 20 μg/mL) for 12 h. CLOCK mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( b ) HaCaT cells were treated as in ( a ) and CLOCK mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. ( c ) HaCaT/shCont and HaCaT/shClock cells were treated with agerarin (20 μg/mL). After 24 h, AQP3 mRNA expression was analyzed by RT-PCR. GAPDH expression was used as an internal control. ( d ) HaCaT cells were treated as in ( c ) and AQP3 mRNA levels were measured by qRT-PCR. GAPDH mRNA level was used for normalization. Full-length gels ( a and c ) are presented in Supplemental Fig. .

    Article Snippet: AQP3 protein was detected using 1:500 diluted ant-AQP3 antibody (BOSTER).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Quantitative RT-PCR