mouse anti pi 3 5 p 2 antibody  (Echelon Biosciences)


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    Echelon Biosciences mouse anti pi 3 5 p 2 antibody
    (A) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. PI(3,5)P 2 (red) was detected by immunofluorescence analysis. Scale bar: 10 μm. (B) Quantification of the relative intensity of PI(3,5)P 2 from (A) (n = 20). (C) PK-15 cells were infected with PRV HN1201 (MOI = 0.1) for 0–24 hours. PIKfyve, MTM1, and PRV gE were detected by immunoblotting. (D) PK-15 cells were mock-infected or infected with PRV HN1201 (MOI = 0.1) and treated with YM-201636 (10 μM) for 24 hours. PI(3,5)P 2 in the lysosomal fraction was detected by lipid dot-blot analysis. The fraction was analyzed by immunoblotting with an antibody against NPC1 (lysosome marker). (E) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. The colocalization of PI(3,5)P 2 (red) with LAMP1 (lysosomal marker, blue) was detected by immunofluorescence analysis. Scale bar: 10 μm. (F) Quantification of colocalization of PI(3,5)P 2 with LAMP1 from (E) (n = 40). (G) PK-15 cells were treated with vehicle, YM-201636 (10 μM), or ML-SI1 (10 μM) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (H) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (I) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were incubated with PRV HN1201 (MOI = 0.1), vehicle, or poly-lysine (50 μg/mL) as indicated at 4°C for 2 hours, and then temperature-shifted to 37°C for 10 minutes to allow entry. After washing with trypsin (1 mg/mL) to remove residual virions on the PM, viral entry was detected by qRT-PCR analysis of viral genome copy numbers in the cells. (J) Schematic representation of the rapamycin-inducible heterodimerization system used to recruit MTM1 to the lysosomal membrane. Rab7 is a lysosome-specific Rab protein. MTM1 is a PI 3 phosphatase that can convert PI(3,5)P 2 and PI(3)P into PI(5)P and PI, respectively. (K) HeLa cells were cotransfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. The subcellular localization of mCherry-FRB-MTM1 and BFP-FKBP-Rab7 was observed by fluorescence microscopy. Scale bar: 10 μm. (L) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. Intracellular Ca 2+ levels were detected using Fluo-3 staining (5 μM). Scale bar: 10 μm. (M) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes and infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (N) Mcoln1 +/+ and Mcoln1 -/- MEF cells were transfected with TRPML1-WT or TRPML1-7Q for 24 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). Data are expressed as the mean ± SD of 3 independent experiments. P -values were determined by Student’s t -test. P < 0.05 was considered statistically significant.
    Mouse Anti Pi 3 5 P 2 Antibody, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti pi 3 5 p 2 antibody/product/Echelon Biosciences
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti pi 3 5 p 2 antibody - by Bioz Stars, 2024-07
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    1) Product Images from "Pseudorabies virus inhibits progesterone-induced inactivation of TRPML1 to facilitate viral entry"

    Article Title: Pseudorabies virus inhibits progesterone-induced inactivation of TRPML1 to facilitate viral entry

    Journal: PLOS Pathogens

    doi: 10.1371/journal.ppat.1011956

    (A) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. PI(3,5)P 2 (red) was detected by immunofluorescence analysis. Scale bar: 10 μm. (B) Quantification of the relative intensity of PI(3,5)P 2 from (A) (n = 20). (C) PK-15 cells were infected with PRV HN1201 (MOI = 0.1) for 0–24 hours. PIKfyve, MTM1, and PRV gE were detected by immunoblotting. (D) PK-15 cells were mock-infected or infected with PRV HN1201 (MOI = 0.1) and treated with YM-201636 (10 μM) for 24 hours. PI(3,5)P 2 in the lysosomal fraction was detected by lipid dot-blot analysis. The fraction was analyzed by immunoblotting with an antibody against NPC1 (lysosome marker). (E) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. The colocalization of PI(3,5)P 2 (red) with LAMP1 (lysosomal marker, blue) was detected by immunofluorescence analysis. Scale bar: 10 μm. (F) Quantification of colocalization of PI(3,5)P 2 with LAMP1 from (E) (n = 40). (G) PK-15 cells were treated with vehicle, YM-201636 (10 μM), or ML-SI1 (10 μM) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (H) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (I) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were incubated with PRV HN1201 (MOI = 0.1), vehicle, or poly-lysine (50 μg/mL) as indicated at 4°C for 2 hours, and then temperature-shifted to 37°C for 10 minutes to allow entry. After washing with trypsin (1 mg/mL) to remove residual virions on the PM, viral entry was detected by qRT-PCR analysis of viral genome copy numbers in the cells. (J) Schematic representation of the rapamycin-inducible heterodimerization system used to recruit MTM1 to the lysosomal membrane. Rab7 is a lysosome-specific Rab protein. MTM1 is a PI 3 phosphatase that can convert PI(3,5)P 2 and PI(3)P into PI(5)P and PI, respectively. (K) HeLa cells were cotransfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. The subcellular localization of mCherry-FRB-MTM1 and BFP-FKBP-Rab7 was observed by fluorescence microscopy. Scale bar: 10 μm. (L) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. Intracellular Ca 2+ levels were detected using Fluo-3 staining (5 μM). Scale bar: 10 μm. (M) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes and infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (N) Mcoln1 +/+ and Mcoln1 -/- MEF cells were transfected with TRPML1-WT or TRPML1-7Q for 24 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). Data are expressed as the mean ± SD of 3 independent experiments. P -values were determined by Student’s t -test. P < 0.05 was considered statistically significant.
    Figure Legend Snippet: (A) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. PI(3,5)P 2 (red) was detected by immunofluorescence analysis. Scale bar: 10 μm. (B) Quantification of the relative intensity of PI(3,5)P 2 from (A) (n = 20). (C) PK-15 cells were infected with PRV HN1201 (MOI = 0.1) for 0–24 hours. PIKfyve, MTM1, and PRV gE were detected by immunoblotting. (D) PK-15 cells were mock-infected or infected with PRV HN1201 (MOI = 0.1) and treated with YM-201636 (10 μM) for 24 hours. PI(3,5)P 2 in the lysosomal fraction was detected by lipid dot-blot analysis. The fraction was analyzed by immunoblotting with an antibody against NPC1 (lysosome marker). (E) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. The colocalization of PI(3,5)P 2 (red) with LAMP1 (lysosomal marker, blue) was detected by immunofluorescence analysis. Scale bar: 10 μm. (F) Quantification of colocalization of PI(3,5)P 2 with LAMP1 from (E) (n = 40). (G) PK-15 cells were treated with vehicle, YM-201636 (10 μM), or ML-SI1 (10 μM) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (H) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (I) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were incubated with PRV HN1201 (MOI = 0.1), vehicle, or poly-lysine (50 μg/mL) as indicated at 4°C for 2 hours, and then temperature-shifted to 37°C for 10 minutes to allow entry. After washing with trypsin (1 mg/mL) to remove residual virions on the PM, viral entry was detected by qRT-PCR analysis of viral genome copy numbers in the cells. (J) Schematic representation of the rapamycin-inducible heterodimerization system used to recruit MTM1 to the lysosomal membrane. Rab7 is a lysosome-specific Rab protein. MTM1 is a PI 3 phosphatase that can convert PI(3,5)P 2 and PI(3)P into PI(5)P and PI, respectively. (K) HeLa cells were cotransfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. The subcellular localization of mCherry-FRB-MTM1 and BFP-FKBP-Rab7 was observed by fluorescence microscopy. Scale bar: 10 μm. (L) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. Intracellular Ca 2+ levels were detected using Fluo-3 staining (5 μM). Scale bar: 10 μm. (M) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes and infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (N) Mcoln1 +/+ and Mcoln1 -/- MEF cells were transfected with TRPML1-WT or TRPML1-7Q for 24 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). Data are expressed as the mean ± SD of 3 independent experiments. P -values were determined by Student’s t -test. P < 0.05 was considered statistically significant.

    Techniques Used: Infection, Immunofluorescence, Western Blot, Dot Blot, Marker, Staining, Incubation, Quantitative RT-PCR, Membrane, Fluorescence, Microscopy, Transfection

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    Echelon Biosciences mouse anti pi 3 5 p 2 antibody
    (A) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. PI(3,5)P 2 (red) was detected by immunofluorescence analysis. Scale bar: 10 μm. (B) Quantification of the relative intensity of PI(3,5)P 2 from (A) (n = 20). (C) PK-15 cells were infected with PRV HN1201 (MOI = 0.1) for 0–24 hours. PIKfyve, MTM1, and PRV gE were detected by immunoblotting. (D) PK-15 cells were mock-infected or infected with PRV HN1201 (MOI = 0.1) and treated with YM-201636 (10 μM) for 24 hours. PI(3,5)P 2 in the lysosomal fraction was detected by lipid dot-blot analysis. The fraction was analyzed by immunoblotting with an antibody against NPC1 (lysosome marker). (E) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. The colocalization of PI(3,5)P 2 (red) with LAMP1 (lysosomal marker, blue) was detected by immunofluorescence analysis. Scale bar: 10 μm. (F) Quantification of colocalization of PI(3,5)P 2 with LAMP1 from (E) (n = 40). (G) PK-15 cells were treated with vehicle, YM-201636 (10 μM), or ML-SI1 (10 μM) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (H) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (I) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were incubated with PRV HN1201 (MOI = 0.1), vehicle, or poly-lysine (50 μg/mL) as indicated at 4°C for 2 hours, and then temperature-shifted to 37°C for 10 minutes to allow entry. After washing with trypsin (1 mg/mL) to remove residual virions on the PM, viral entry was detected by qRT-PCR analysis of viral genome copy numbers in the cells. (J) Schematic representation of the rapamycin-inducible heterodimerization system used to recruit MTM1 to the lysosomal membrane. Rab7 is a lysosome-specific Rab protein. MTM1 is a PI 3 phosphatase that can convert PI(3,5)P 2 and PI(3)P into PI(5)P and PI, respectively. (K) HeLa cells were cotransfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. The subcellular localization of mCherry-FRB-MTM1 and BFP-FKBP-Rab7 was observed by fluorescence microscopy. Scale bar: 10 μm. (L) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. Intracellular Ca 2+ levels were detected using Fluo-3 staining (5 μM). Scale bar: 10 μm. (M) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes and infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (N) Mcoln1 +/+ and Mcoln1 -/- MEF cells were transfected with TRPML1-WT or TRPML1-7Q for 24 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). Data are expressed as the mean ± SD of 3 independent experiments. P -values were determined by Student’s t -test. P < 0.05 was considered statistically significant.
    Mouse Anti Pi 3 5 P 2 Antibody, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti pi 3 5 p 2 antibody/product/Echelon Biosciences
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti pi 3 5 p 2 antibody - by Bioz Stars, 2024-07
    86/100 stars
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    (A) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. PI(3,5)P 2 (red) was detected by immunofluorescence analysis. Scale bar: 10 μm. (B) Quantification of the relative intensity of PI(3,5)P 2 from (A) (n = 20). (C) PK-15 cells were infected with PRV HN1201 (MOI = 0.1) for 0–24 hours. PIKfyve, MTM1, and PRV gE were detected by immunoblotting. (D) PK-15 cells were mock-infected or infected with PRV HN1201 (MOI = 0.1) and treated with YM-201636 (10 μM) for 24 hours. PI(3,5)P 2 in the lysosomal fraction was detected by lipid dot-blot analysis. The fraction was analyzed by immunoblotting with an antibody against NPC1 (lysosome marker). (E) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. The colocalization of PI(3,5)P 2 (red) with LAMP1 (lysosomal marker, blue) was detected by immunofluorescence analysis. Scale bar: 10 μm. (F) Quantification of colocalization of PI(3,5)P 2 with LAMP1 from (E) (n = 40). (G) PK-15 cells were treated with vehicle, YM-201636 (10 μM), or ML-SI1 (10 μM) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (H) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (I) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were incubated with PRV HN1201 (MOI = 0.1), vehicle, or poly-lysine (50 μg/mL) as indicated at 4°C for 2 hours, and then temperature-shifted to 37°C for 10 minutes to allow entry. After washing with trypsin (1 mg/mL) to remove residual virions on the PM, viral entry was detected by qRT-PCR analysis of viral genome copy numbers in the cells. (J) Schematic representation of the rapamycin-inducible heterodimerization system used to recruit MTM1 to the lysosomal membrane. Rab7 is a lysosome-specific Rab protein. MTM1 is a PI 3 phosphatase that can convert PI(3,5)P 2 and PI(3)P into PI(5)P and PI, respectively. (K) HeLa cells were cotransfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. The subcellular localization of mCherry-FRB-MTM1 and BFP-FKBP-Rab7 was observed by fluorescence microscopy. Scale bar: 10 μm. (L) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. Intracellular Ca 2+ levels were detected using Fluo-3 staining (5 μM). Scale bar: 10 μm. (M) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes and infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (N) Mcoln1 +/+ and Mcoln1 -/- MEF cells were transfected with TRPML1-WT or TRPML1-7Q for 24 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). Data are expressed as the mean ± SD of 3 independent experiments. P -values were determined by Student’s t -test. P < 0.05 was considered statistically significant.

    Journal: PLOS Pathogens

    Article Title: Pseudorabies virus inhibits progesterone-induced inactivation of TRPML1 to facilitate viral entry

    doi: 10.1371/journal.ppat.1011956

    Figure Lengend Snippet: (A) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. PI(3,5)P 2 (red) was detected by immunofluorescence analysis. Scale bar: 10 μm. (B) Quantification of the relative intensity of PI(3,5)P 2 from (A) (n = 20). (C) PK-15 cells were infected with PRV HN1201 (MOI = 0.1) for 0–24 hours. PIKfyve, MTM1, and PRV gE were detected by immunoblotting. (D) PK-15 cells were mock-infected or infected with PRV HN1201 (MOI = 0.1) and treated with YM-201636 (10 μM) for 24 hours. PI(3,5)P 2 in the lysosomal fraction was detected by lipid dot-blot analysis. The fraction was analyzed by immunoblotting with an antibody against NPC1 (lysosome marker). (E) PK-15 cells were mock-infected or infected with PRV-GFP (MOI = 0.1) for 24 hours. The colocalization of PI(3,5)P 2 (red) with LAMP1 (lysosomal marker, blue) was detected by immunofluorescence analysis. Scale bar: 10 μm. (F) Quantification of colocalization of PI(3,5)P 2 with LAMP1 from (E) (n = 40). (G) PK-15 cells were treated with vehicle, YM-201636 (10 μM), or ML-SI1 (10 μM) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (H) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (I) PK-15 cells were treated with vehicle or poly-lysine (50 μg/mL) for 8 hours. Cells were incubated with PRV HN1201 (MOI = 0.1), vehicle, or poly-lysine (50 μg/mL) as indicated at 4°C for 2 hours, and then temperature-shifted to 37°C for 10 minutes to allow entry. After washing with trypsin (1 mg/mL) to remove residual virions on the PM, viral entry was detected by qRT-PCR analysis of viral genome copy numbers in the cells. (J) Schematic representation of the rapamycin-inducible heterodimerization system used to recruit MTM1 to the lysosomal membrane. Rab7 is a lysosome-specific Rab protein. MTM1 is a PI 3 phosphatase that can convert PI(3,5)P 2 and PI(3)P into PI(5)P and PI, respectively. (K) HeLa cells were cotransfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. The subcellular localization of mCherry-FRB-MTM1 and BFP-FKBP-Rab7 was observed by fluorescence microscopy. Scale bar: 10 μm. (L) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes. Intracellular Ca 2+ levels were detected using Fluo-3 staining (5 μM). Scale bar: 10 μm. (M) HeLa cells were transfected with mCherry-FRB-MTM1 (red) and BFP-FKBP-Rab7 (blue) for 24 hours. Cells were then treated with rapamycin (500 nM) for 20 minutes and infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). (N) Mcoln1 +/+ and Mcoln1 -/- MEF cells were transfected with TRPML1-WT or TRPML1-7Q for 24 hours. Cells were then infected with PRV HN1201 (MOI = 0.1). Intracellular Ca 2+ levels were immediately detected using Fluo-3 staining (5 μM). Data are expressed as the mean ± SD of 3 independent experiments. P -values were determined by Student’s t -test. P < 0.05 was considered statistically significant.

    Article Snippet: The membrane was then blocked in 3% fatty acid-free BSA at room temperature for 1 h and probed with mouse anti-PI(3,5)P 2 antibody (1:1,000 dilution; Echelon).

    Techniques: Infection, Immunofluorescence, Western Blot, Dot Blot, Marker, Staining, Incubation, Quantitative RT-PCR, Membrane, Fluorescence, Microscopy, Transfection