gabaar subunit α1  (Alomone Labs)


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    Structured Review

    Alomone Labs gabaar subunit α1
    High magnification (63X) images obtained with laser scanning confocal microscopy reveal the diffuse distribution of <t>GABAAR</t> <t>α1,</t> β2/3, and γ2 subunits and GAD65 immunoreactivity in control MD. The distribution of IR of α1 (A), β2/3 (B), and γ2 (C) subunits was similar and found to be present as many small diffuse clusters and some large clusters. The immunoreactivity of GAD65 (D) was mainly found as large intense clusters. The scale bar in figure A is 7 µm.
    Gabaar Subunit α1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gabaar subunit α1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gabaar subunit α1 - by Bioz Stars, 2022-07
    93/100 stars

    Images

    1) Product Images from "Altered pharmacology and GABA-A receptor subunit expression in dorsal midline thalamic neurons in limbic epilepsy"

    Article Title: Altered pharmacology and GABA-A receptor subunit expression in dorsal midline thalamic neurons in limbic epilepsy

    Journal: Neurobiology of disease

    doi: 10.1016/j.nbd.2008.09.023

    High magnification (63X) images obtained with laser scanning confocal microscopy reveal the diffuse distribution of GABAAR α1, β2/3, and γ2 subunits and GAD65 immunoreactivity in control MD. The distribution of IR of α1 (A), β2/3 (B), and γ2 (C) subunits was similar and found to be present as many small diffuse clusters and some large clusters. The immunoreactivity of GAD65 (D) was mainly found as large intense clusters. The scale bar in figure A is 7 µm.
    Figure Legend Snippet: High magnification (63X) images obtained with laser scanning confocal microscopy reveal the diffuse distribution of GABAAR α1, β2/3, and γ2 subunits and GAD65 immunoreactivity in control MD. The distribution of IR of α1 (A), β2/3 (B), and γ2 (C) subunits was similar and found to be present as many small diffuse clusters and some large clusters. The immunoreactivity of GAD65 (D) was mainly found as large intense clusters. The scale bar in figure A is 7 µm.

    Techniques Used: Confocal Microscopy

    Distribution of α1, β2/3, γ2, of the GABAAR, and GAD-IR in thalamus captured at low magnification (2x, NA = 0.1) to demonstrate different intensity of staining at different areas. The white arrows point to PV and the white arrowheads point to MD. The α1, β2/3, γ2, and GAD-IR were expressed in most thalamic nuclei, and the entire thalamus appeared uniformly stained for these subunits, except for the reticular nucleus and the habenula or hippocampus (A–D). The staining was diffuse, and individual neurons could not be readily distinguished, except that of α1-IR in hippocampal dentate gyrus (A). The scale bar in figure A is 200 µm, and also applies to B, C and D.
    Figure Legend Snippet: Distribution of α1, β2/3, γ2, of the GABAAR, and GAD-IR in thalamus captured at low magnification (2x, NA = 0.1) to demonstrate different intensity of staining at different areas. The white arrows point to PV and the white arrowheads point to MD. The α1, β2/3, γ2, and GAD-IR were expressed in most thalamic nuclei, and the entire thalamus appeared uniformly stained for these subunits, except for the reticular nucleus and the habenula or hippocampus (A–D). The staining was diffuse, and individual neurons could not be readily distinguished, except that of α1-IR in hippocampal dentate gyrus (A). The scale bar in figure A is 200 µm, and also applies to B, C and D.

    Techniques Used: Staining

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    Alomone Labs gabaar subunit α1
    High magnification (63X) images obtained with laser scanning confocal microscopy reveal the diffuse distribution of <t>GABAAR</t> <t>α1,</t> β2/3, and γ2 subunits and GAD65 immunoreactivity in control MD. The distribution of IR of α1 (A), β2/3 (B), and γ2 (C) subunits was similar and found to be present as many small diffuse clusters and some large clusters. The immunoreactivity of GAD65 (D) was mainly found as large intense clusters. The scale bar in figure A is 7 µm.
    Gabaar Subunit α1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gabaar subunit α1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gabaar subunit α1 - by Bioz Stars, 2022-07
    93/100 stars
      Buy from Supplier

    91
    Alomone Labs anti mcoln3 trpml3
    Effect of palmitoylation on <t>MCOLN3/TRPML3</t> surface expression. (a) Analysis of surface expression by biotinylation. HEK293T cells expressing WT MCOLN3/TRPML3, MCOLN3/TRPML3(C11S), or MCOLN3/TRPML3(3CS) treated with ethanol (control) or 2-BP, and HeLa cells expressing WT MCOLN3/TRPML3 or MCOLN3/TRPML3(3CS) were used for surface biotinylation assays. Total and surface proteins were analyzed by western blot using anti-GFP antibody. The data are representative of 3 independent experiments. (b) Densitometric analysis of surface MCOLN3/TRPML3s to total MCOLN3/TRPML3 protein levels (n = 3, * p
    Anti Mcoln3 Trpml3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mcoln3 trpml3/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti mcoln3 trpml3 - by Bioz Stars, 2022-07
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    Image Search Results


    High magnification (63X) images obtained with laser scanning confocal microscopy reveal the diffuse distribution of GABAAR α1, β2/3, and γ2 subunits and GAD65 immunoreactivity in control MD. The distribution of IR of α1 (A), β2/3 (B), and γ2 (C) subunits was similar and found to be present as many small diffuse clusters and some large clusters. The immunoreactivity of GAD65 (D) was mainly found as large intense clusters. The scale bar in figure A is 7 µm.

    Journal: Neurobiology of disease

    Article Title: Altered pharmacology and GABA-A receptor subunit expression in dorsal midline thalamic neurons in limbic epilepsy

    doi: 10.1016/j.nbd.2008.09.023

    Figure Lengend Snippet: High magnification (63X) images obtained with laser scanning confocal microscopy reveal the diffuse distribution of GABAAR α1, β2/3, and γ2 subunits and GAD65 immunoreactivity in control MD. The distribution of IR of α1 (A), β2/3 (B), and γ2 (C) subunits was similar and found to be present as many small diffuse clusters and some large clusters. The immunoreactivity of GAD65 (D) was mainly found as large intense clusters. The scale bar in figure A is 7 µm.

    Article Snippet: The antibodies against the GABAAR subunit α1 (1–16) and γ2 (1–33) were obtained from Alomone labs (Israel).

    Techniques: Confocal Microscopy

    Distribution of α1, β2/3, γ2, of the GABAAR, and GAD-IR in thalamus captured at low magnification (2x, NA = 0.1) to demonstrate different intensity of staining at different areas. The white arrows point to PV and the white arrowheads point to MD. The α1, β2/3, γ2, and GAD-IR were expressed in most thalamic nuclei, and the entire thalamus appeared uniformly stained for these subunits, except for the reticular nucleus and the habenula or hippocampus (A–D). The staining was diffuse, and individual neurons could not be readily distinguished, except that of α1-IR in hippocampal dentate gyrus (A). The scale bar in figure A is 200 µm, and also applies to B, C and D.

    Journal: Neurobiology of disease

    Article Title: Altered pharmacology and GABA-A receptor subunit expression in dorsal midline thalamic neurons in limbic epilepsy

    doi: 10.1016/j.nbd.2008.09.023

    Figure Lengend Snippet: Distribution of α1, β2/3, γ2, of the GABAAR, and GAD-IR in thalamus captured at low magnification (2x, NA = 0.1) to demonstrate different intensity of staining at different areas. The white arrows point to PV and the white arrowheads point to MD. The α1, β2/3, γ2, and GAD-IR were expressed in most thalamic nuclei, and the entire thalamus appeared uniformly stained for these subunits, except for the reticular nucleus and the habenula or hippocampus (A–D). The staining was diffuse, and individual neurons could not be readily distinguished, except that of α1-IR in hippocampal dentate gyrus (A). The scale bar in figure A is 200 µm, and also applies to B, C and D.

    Article Snippet: The antibodies against the GABAAR subunit α1 (1–16) and γ2 (1–33) were obtained from Alomone labs (Israel).

    Techniques: Staining

    Effect of palmitoylation on MCOLN3/TRPML3 surface expression. (a) Analysis of surface expression by biotinylation. HEK293T cells expressing WT MCOLN3/TRPML3, MCOLN3/TRPML3(C11S), or MCOLN3/TRPML3(3CS) treated with ethanol (control) or 2-BP, and HeLa cells expressing WT MCOLN3/TRPML3 or MCOLN3/TRPML3(3CS) were used for surface biotinylation assays. Total and surface proteins were analyzed by western blot using anti-GFP antibody. The data are representative of 3 independent experiments. (b) Densitometric analysis of surface MCOLN3/TRPML3s to total MCOLN3/TRPML3 protein levels (n = 3, * p

    Journal: Autophagy

    Article Title: Palmitoylation controls trafficking of the intracellular Ca2+ channel MCOLN3/TRPML3 to regulate autophagy

    doi: 10.1080/15548627.2018.1518671

    Figure Lengend Snippet: Effect of palmitoylation on MCOLN3/TRPML3 surface expression. (a) Analysis of surface expression by biotinylation. HEK293T cells expressing WT MCOLN3/TRPML3, MCOLN3/TRPML3(C11S), or MCOLN3/TRPML3(3CS) treated with ethanol (control) or 2-BP, and HeLa cells expressing WT MCOLN3/TRPML3 or MCOLN3/TRPML3(3CS) were used for surface biotinylation assays. Total and surface proteins were analyzed by western blot using anti-GFP antibody. The data are representative of 3 independent experiments. (b) Densitometric analysis of surface MCOLN3/TRPML3s to total MCOLN3/TRPML3 protein levels (n = 3, * p

    Article Snippet: The following antibodies were used: anti-LAMP1 (BD biosciences, 555798), anti-GFP (Invitrogen, ), anti-LC3 (Thermo Fisher Scientific, PA1-16930), and anti-MCOLN3/TRPML3 (Alomone Labs, ACC-083).

    Techniques: Expressing, Western Blot

    Effect of palmitoylation on MCOLN3/TRPML3 trafficking in the context of autophagy. ( A-C ) HeLa cells transfected with a plasmid encoding GFP-LC3 only (a) or co-transfected with a plasmid encoding mCherry-MCOLN3/TRPML3 or mCherry-MCOLN3/TRPML3(3CS) were kept in fed medium (b) or serum starved for 2 h (c). (d and e) The number of GFP-LC3 puncta in serum-starved cells (d) or in cells treated with 25 µM CPA for 3 h (e) was counted using ImageJ and given as the mean ± SEM of 12–28 cells (* p

    Journal: Autophagy

    Article Title: Palmitoylation controls trafficking of the intracellular Ca2+ channel MCOLN3/TRPML3 to regulate autophagy

    doi: 10.1080/15548627.2018.1518671

    Figure Lengend Snippet: Effect of palmitoylation on MCOLN3/TRPML3 trafficking in the context of autophagy. ( A-C ) HeLa cells transfected with a plasmid encoding GFP-LC3 only (a) or co-transfected with a plasmid encoding mCherry-MCOLN3/TRPML3 or mCherry-MCOLN3/TRPML3(3CS) were kept in fed medium (b) or serum starved for 2 h (c). (d and e) The number of GFP-LC3 puncta in serum-starved cells (d) or in cells treated with 25 µM CPA for 3 h (e) was counted using ImageJ and given as the mean ± SEM of 12–28 cells (* p

    Article Snippet: The following antibodies were used: anti-LAMP1 (BD biosciences, 555798), anti-GFP (Invitrogen, ), anti-LC3 (Thermo Fisher Scientific, PA1-16930), and anti-MCOLN3/TRPML3 (Alomone Labs, ACC-083).

    Techniques: Transfection, Plasmid Preparation

    Organellar localization and Ca 2+ release by WT MCOLN3/TRPML3 and MCOLN3/TRPML3(3CS). (a) HeLa cells transfected with plasmids encoding mCherry-MCOLN3/TRPML3 or mCherry-MCOLN3/TRPML3(3CS) were stained for LAMP1 (green) and imaged by confocal microscopy. (b) Images similar to those in panel (a) from 5–9 cells were used to determine the overlap with ImageJ. The results are given as the mean ± SEM. (c) HeLa cells transfected with a plasmid encoding mCherry-MCOLN3/TRPML3 or mCherry-MCOLN3/TRPML3(3CS) and GFP-LC3 were imaged by confocal microscopy. (d) Images similar to those in panel (c) from 7 cells were used to determine the overlap with ImageJ. The results are given as the mean ± SEM. (e) HEK293T cells expressing GFP (gray trace), WT MCOLN3/TRPML3 (black trace) or MCOLN3/TRPML3(3CS) (red trace) were preincubated in Ca 2+ -free medium to block PM Ca 2+ influx. Organellar Ca 2+ efflux was measured by applying 20 µM ML-SA1 in Ca 2+ -free medium and then, Ca 2+ influx from the PM was measured by increasing Ca 2+ o to 10 mM. (f) Cellular responses to ML-SA1 and 10 mM Ca 2+ o were compared in cells transfected with WT MCOLN3/TRPML3 and MCOLN3/TRPML3(3CS). Results are plotted as the mean ± SEM of 28–29 cells (*** p

    Journal: Autophagy

    Article Title: Palmitoylation controls trafficking of the intracellular Ca2+ channel MCOLN3/TRPML3 to regulate autophagy

    doi: 10.1080/15548627.2018.1518671

    Figure Lengend Snippet: Organellar localization and Ca 2+ release by WT MCOLN3/TRPML3 and MCOLN3/TRPML3(3CS). (a) HeLa cells transfected with plasmids encoding mCherry-MCOLN3/TRPML3 or mCherry-MCOLN3/TRPML3(3CS) were stained for LAMP1 (green) and imaged by confocal microscopy. (b) Images similar to those in panel (a) from 5–9 cells were used to determine the overlap with ImageJ. The results are given as the mean ± SEM. (c) HeLa cells transfected with a plasmid encoding mCherry-MCOLN3/TRPML3 or mCherry-MCOLN3/TRPML3(3CS) and GFP-LC3 were imaged by confocal microscopy. (d) Images similar to those in panel (c) from 7 cells were used to determine the overlap with ImageJ. The results are given as the mean ± SEM. (e) HEK293T cells expressing GFP (gray trace), WT MCOLN3/TRPML3 (black trace) or MCOLN3/TRPML3(3CS) (red trace) were preincubated in Ca 2+ -free medium to block PM Ca 2+ influx. Organellar Ca 2+ efflux was measured by applying 20 µM ML-SA1 in Ca 2+ -free medium and then, Ca 2+ influx from the PM was measured by increasing Ca 2+ o to 10 mM. (f) Cellular responses to ML-SA1 and 10 mM Ca 2+ o were compared in cells transfected with WT MCOLN3/TRPML3 and MCOLN3/TRPML3(3CS). Results are plotted as the mean ± SEM of 28–29 cells (*** p

    Article Snippet: The following antibodies were used: anti-LAMP1 (BD biosciences, 555798), anti-GFP (Invitrogen, ), anti-LC3 (Thermo Fisher Scientific, PA1-16930), and anti-MCOLN3/TRPML3 (Alomone Labs, ACC-083).

    Techniques: Transfection, Staining, Confocal Microscopy, Plasmid Preparation, Expressing, Blocking Assay

    Palmitoylation sites and PATs of MCOLN3/TRPML3. (a) Schematic of MCOLN3/TRPML3 showing palmitoylation sites (gray and red circles) predicted by CSS-Palm v2.0 and sequence alignments of the indicated cysteine residues across different species and among MCOLN/TRPML subfamily members. (b) MCOLN3/TRPML3 palmitoylation assay. HEK293T cells expressing GFP-tagged MCOLN3/TRPML3 (upper panel) or HEK293T and SW13 cells (lower panel) were either treated with DMSO (control) or metabolically labeled with 100 μM 17-ODYA. After 6 h, membrane fractions were prepared followed by reaction between biotin-azide and 17-ODYA using click chemistry. Biotinylated proteins were affinity isolated using streptavidin beads and eluates were analyzed by western blot using anti-GFP antibody or anti-MCOLN3/TRPML3 antibody to detect palmitoylated proteins. Tris or HN were added to the reaction mixture and incubated for 1 h at RT before adding the beads. Input samples were 1% of protein lysate used in the click chemistry reactions. (c) MCOLN3/TRPML3 and cysteine mutant palmitoylation assays. HEK293T cells expressing GFP-tagged MCOLN3/TRPML3 and the indicated single or triple cysteine mutants were assayed as in (b). (d) Quantified data of ratios of palmitoylated MCOLN3/TRPML3s to total MCOLN3/TRPML3 protein levels (n = 3, * p

    Journal: Autophagy

    Article Title: Palmitoylation controls trafficking of the intracellular Ca2+ channel MCOLN3/TRPML3 to regulate autophagy

    doi: 10.1080/15548627.2018.1518671

    Figure Lengend Snippet: Palmitoylation sites and PATs of MCOLN3/TRPML3. (a) Schematic of MCOLN3/TRPML3 showing palmitoylation sites (gray and red circles) predicted by CSS-Palm v2.0 and sequence alignments of the indicated cysteine residues across different species and among MCOLN/TRPML subfamily members. (b) MCOLN3/TRPML3 palmitoylation assay. HEK293T cells expressing GFP-tagged MCOLN3/TRPML3 (upper panel) or HEK293T and SW13 cells (lower panel) were either treated with DMSO (control) or metabolically labeled with 100 μM 17-ODYA. After 6 h, membrane fractions were prepared followed by reaction between biotin-azide and 17-ODYA using click chemistry. Biotinylated proteins were affinity isolated using streptavidin beads and eluates were analyzed by western blot using anti-GFP antibody or anti-MCOLN3/TRPML3 antibody to detect palmitoylated proteins. Tris or HN were added to the reaction mixture and incubated for 1 h at RT before adding the beads. Input samples were 1% of protein lysate used in the click chemistry reactions. (c) MCOLN3/TRPML3 and cysteine mutant palmitoylation assays. HEK293T cells expressing GFP-tagged MCOLN3/TRPML3 and the indicated single or triple cysteine mutants were assayed as in (b). (d) Quantified data of ratios of palmitoylated MCOLN3/TRPML3s to total MCOLN3/TRPML3 protein levels (n = 3, * p

    Article Snippet: The following antibodies were used: anti-LAMP1 (BD biosciences, 555798), anti-GFP (Invitrogen, ), anti-LC3 (Thermo Fisher Scientific, PA1-16930), and anti-MCOLN3/TRPML3 (Alomone Labs, ACC-083).

    Techniques: Sequencing, Expressing, Metabolic Labelling, Labeling, Isolation, Western Blot, Incubation, Mutagenesis

    Effects of palmitoylation on MCOLN3/TRPML3 trafficking. (a) Schematic for MCOLN3/TRPML3 trafficking to and from the PM. (b and d) HEK293T cells expressing WT MCOLN3/TRPML3 or MCOLN3/TRPML3(3CS) were treated with either 20 µM BFA (b) for 4 h or 100 µM Dynasore (d) for 2 h at 37°C. MCOLN3/TRPML3 currents were measured by applying 5 µM SF21 in 140 mM Na + bath solution. I/V relationships of the MCOLN3/TRPML3 current recorded from cells treated with the indicated compound (black and red traces) at the indicated time points are shown. (c and e) The current amplitudes and relative current density of WT MCOLN3/TRPML3 (black circle) and MCOLN3/TRPML3(3CS) (red circle) in experiments similar to those in panel (b and d) were plotted as the mean ± SEM of 10–21 cells (* p

    Journal: Autophagy

    Article Title: Palmitoylation controls trafficking of the intracellular Ca2+ channel MCOLN3/TRPML3 to regulate autophagy

    doi: 10.1080/15548627.2018.1518671

    Figure Lengend Snippet: Effects of palmitoylation on MCOLN3/TRPML3 trafficking. (a) Schematic for MCOLN3/TRPML3 trafficking to and from the PM. (b and d) HEK293T cells expressing WT MCOLN3/TRPML3 or MCOLN3/TRPML3(3CS) were treated with either 20 µM BFA (b) for 4 h or 100 µM Dynasore (d) for 2 h at 37°C. MCOLN3/TRPML3 currents were measured by applying 5 µM SF21 in 140 mM Na + bath solution. I/V relationships of the MCOLN3/TRPML3 current recorded from cells treated with the indicated compound (black and red traces) at the indicated time points are shown. (c and e) The current amplitudes and relative current density of WT MCOLN3/TRPML3 (black circle) and MCOLN3/TRPML3(3CS) (red circle) in experiments similar to those in panel (b and d) were plotted as the mean ± SEM of 10–21 cells (* p

    Article Snippet: The following antibodies were used: anti-LAMP1 (BD biosciences, 555798), anti-GFP (Invitrogen, ), anti-LC3 (Thermo Fisher Scientific, PA1-16930), and anti-MCOLN3/TRPML3 (Alomone Labs, ACC-083).

    Techniques: Expressing

    Differential effect of organellar Ca 2+ efflux via WT MCOLN3/TRPML3 and MCOLN3/TRPML3(3CS) on autophagy. (a and b) HeLa cells transfected with a plasmid encoding GFP-LC3 only (a) or co-transfected with a plasmid encoding mCherry-MCOLN3/TRPML3 (b) were treated with vehicle or 20 µM ML-SA1 in Ca 2+ -free medium for 1 h at 37°C. (c) HEK293T cells transfected with a plasmid encoding WT MCOLN3/TRPML3 were preincubated with 10 µM BAPTA-AM for 30 min and intracellular Ca 2+ . (d) HeLa cells expressing GFP-LC3 and mCherry-MCOLN3/TRPML3 were preincubated with 10 µM BAPTA-AM in Ca 2+ -free medium for 30 min and treated as in panel (b). (e) Ca 2+ was measured from HEK293T cells transfected with WT MCOLN3/TRPML3 by applying 500 µM GPN and then 20 µM ML-SA1. (f) HeLa cells expressing GFP-LC3 and mCherry-MCOLN3/TRPML3 were preincubated with 500 µM GPN and 10 µM CPA in Ca 2+ -free medium for 30 min and treated as in panel (b). (g) HeLa cells transfected with a plasmid encoding GFP-LC3 and mCherry-MCOLN3/TRPML3(3CS) were treated as in panel (b). (h) The normalized number of GFP-LC3 puncta in panel (a, b, d, f, g) was counted using ImageJ and given as the mean ± SEM of 23–33 cells for panel (a, b, g) and that of 4–14 cells for panel ( D and F ) (*** p

    Journal: Autophagy

    Article Title: Palmitoylation controls trafficking of the intracellular Ca2+ channel MCOLN3/TRPML3 to regulate autophagy

    doi: 10.1080/15548627.2018.1518671

    Figure Lengend Snippet: Differential effect of organellar Ca 2+ efflux via WT MCOLN3/TRPML3 and MCOLN3/TRPML3(3CS) on autophagy. (a and b) HeLa cells transfected with a plasmid encoding GFP-LC3 only (a) or co-transfected with a plasmid encoding mCherry-MCOLN3/TRPML3 (b) were treated with vehicle or 20 µM ML-SA1 in Ca 2+ -free medium for 1 h at 37°C. (c) HEK293T cells transfected with a plasmid encoding WT MCOLN3/TRPML3 were preincubated with 10 µM BAPTA-AM for 30 min and intracellular Ca 2+ . (d) HeLa cells expressing GFP-LC3 and mCherry-MCOLN3/TRPML3 were preincubated with 10 µM BAPTA-AM in Ca 2+ -free medium for 30 min and treated as in panel (b). (e) Ca 2+ was measured from HEK293T cells transfected with WT MCOLN3/TRPML3 by applying 500 µM GPN and then 20 µM ML-SA1. (f) HeLa cells expressing GFP-LC3 and mCherry-MCOLN3/TRPML3 were preincubated with 500 µM GPN and 10 µM CPA in Ca 2+ -free medium for 30 min and treated as in panel (b). (g) HeLa cells transfected with a plasmid encoding GFP-LC3 and mCherry-MCOLN3/TRPML3(3CS) were treated as in panel (b). (h) The normalized number of GFP-LC3 puncta in panel (a, b, d, f, g) was counted using ImageJ and given as the mean ± SEM of 23–33 cells for panel (a, b, g) and that of 4–14 cells for panel ( D and F ) (*** p

    Article Snippet: The following antibodies were used: anti-LAMP1 (BD biosciences, 555798), anti-GFP (Invitrogen, ), anti-LC3 (Thermo Fisher Scientific, PA1-16930), and anti-MCOLN3/TRPML3 (Alomone Labs, ACC-083).

    Techniques: Transfection, Plasmid Preparation, Expressing