Journal:

Article Title: Global Expression Profiling and Physiological Characterization of Corynebacterium glutamicum Grown in the Presence of l -Valine

doi: 10.1128/AEM.69.5.2521-2532.2003

Figure Lengend Snippet: ORFs showing altered relative mRNA levels in response to l -valine in the valine production strain VAL1 a

Article Snippet: Proteins showing altered abundances on 2-D gels in response to l -valine in the wild-type C. glutamicum ATCC 13032 table ft1 table-wrap mode="anchored" t5 TABLE 5. caption a7 ORF NCBI Annotation Gene Intensity b −Val +Val Ratio 1504 NCgl0899 Similarity to 2-nitropropane dioxygenase ( Williopsis mrakii ) 0.4 0.2 0.5 3194 NCgl2167 Pyruvate dehydrogenase component E1 aceE ( C. glutamicum ) 0.6 0.2 0.3 990 NCgl0478 Strong similarity to translation elongation factor EF-G fusA ( E. coli ) 0.3 0.6 2.3 1414 NCgl0827 Strong similarity to PurH bifunctional enzyme ( E. coli ) 0.3 1.5 4.3 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 1.6 4.9 3.1 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.8 3.1 3.6 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.4 1.5 4.1 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.2 0.7 4.2 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.5 2.4 5.1 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.6 3.4 5.3 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.2 1.0 5.9 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.2 1.6 7.2 Open in a separate window a Abbreviations, conditions, and statistical analysis are described in footnotes to Table . b +Val, growth under addition of 40 mM l -valine.

Techniques:

Journal:

Article Title: Global Expression Profiling and Physiological Characterization of Corynebacterium glutamicum Grown in the Presence of l -Valine

doi: 10.1128/AEM.69.5.2521-2532.2003

Figure Lengend Snippet: Proteins showing altered abundances on 2-D gels in response to l -valine in the valine production strain VAL1 a

Article Snippet: Proteins showing altered abundances on 2-D gels in response to l -valine in the wild-type C. glutamicum ATCC 13032 table ft1 table-wrap mode="anchored" t5 TABLE 5. caption a7 ORF NCBI Annotation Gene Intensity b −Val +Val Ratio 1504 NCgl0899 Similarity to 2-nitropropane dioxygenase ( Williopsis mrakii ) 0.4 0.2 0.5 3194 NCgl2167 Pyruvate dehydrogenase component E1 aceE ( C. glutamicum ) 0.6 0.2 0.3 990 NCgl0478 Strong similarity to translation elongation factor EF-G fusA ( E. coli ) 0.3 0.6 2.3 1414 NCgl0827 Strong similarity to PurH bifunctional enzyme ( E. coli ) 0.3 1.5 4.3 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 1.6 4.9 3.1 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.8 3.1 3.6 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.4 1.5 4.1 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.2 0.7 4.2 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.5 2.4 5.1 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.6 3.4 5.3 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.2 1.0 5.9 2793 NCgl1222 Acetolactate synthase large-chain ilvB ( C. glutamicum ) ilvB 0.2 1.6 7.2 Open in a separate window a Abbreviations, conditions, and statistical analysis are described in footnotes to Table . b +Val, growth under addition of 40 mM l -valine.

Techniques:

Journal: Nature

Article Title: Structural basis of catalytic activation in human splicing.

doi: 10.1038/s41586-023-06049-w

Figure Lengend Snippet: Fig. 3 | PRP2 tightly regulates the dissociation of the RES complex and remodels the SF3B complex. a, Relocation of PRP2 on the intron, RES complex dissociation from the translocated RNA and subsequent binding of PPIL4 to the latter. Pink, green and broken lines represent the PPT, the PPT region bound by SF3B1 within Bact, and the PPT-equivalent region visible only within the budding yeast Bact structure, respectively. b, Close-up view of the interfaces between

Article Snippet: Human PRP2 (NCBI Reference Sequence identifier: NM_003587, transcript variant 1) and human PPIL4 (NCBI Sequence identifiers BC020986 and NM_139126.4) open reading frame clones were obtained from OriGene (RC202912) and Applied Biological Materials (373710120000), whereas human GPKOW and SKIP were codon-optimized for expression in insect cells and synthesized by GeneArt (ThermoFisher Scientific).

Techniques: Binding Assay

Journal: Nature Communications

Article Title: Contribution of GATA6 to homeostasis of the human upper pilosebaceous unit and acne pathogenesis

doi: 10.1038/s41467-020-18784-z

Figure Lengend Snippet: a Schematic of the pilosebaceous unit indicating the upper region, consisting of interfollicular epidermis (IFE), infundibulum (INF), junctional zone (JZ), sebaceous duct (SD), and sebaceous gland (SG), then the isthmus (IST) and the lower region of the hair follicle (HF) comprising the bulge (Bu), hair shaft (HS), inner root sheath (IRS), and outer root sheath (ORS). The lower right panel shows the structure and differentiation of the human SG. b – e Immunolabelling for GATA6 ( b – e ) and ITGα6 ( b ), IVL ( c ), FASN ( d ) and KRT7 ( e ), with DAPI counterstain of back skin sections from a 60 year old male. Middle and right panels in b are higher magnification views of the left panel section. Yellow asterisks show basal GATA6+ cells. Scale bar: 100 μm, except in b right panels: 25 μm. Data are representative of two independent experiments. f 2D Uniform Manifold Approximation and Projection (UMAP) visualisation of single cells isolated from healthy human scalp epidermis and adnexa coloured by Seurat cluster. g GATA6 expression levels, as log 10 Transcripts Per Million (TPM), across the 24 different clusters. GATA6 is mainly expressed in cluster 22. h Differentially expressed genes (DEG) of cluster 22. Only DEG with log 2 FC > 0.2 or < −0.2 are shown. i Intersection of the 250 most upregulated DEG of cluster 22 identified in scalp epidermis within cell populations identified by Cheng et al. HR-WNTI stands for High Resolution-WNT Inhibitory cluster and UHF for upper hair follicle cluster. j GO enrichment analysis of the 250 most upregulated DEG of cluster 22.

Article Snippet: Human GATA6 open reading frame (ORF) (NCBI reference sequence NM_005257.5) cloned into a pReceiver-M02 expression plasmid was purchased from GeneCopoeia (Tebu-Bio).

Techniques: Isolation, Expressing

Journal: Nature Communications

Article Title: Contribution of GATA6 to homeostasis of the human upper pilosebaceous unit and acne pathogenesis

doi: 10.1038/s41467-020-18784-z

Figure Lengend Snippet: a Skin sections from two healthy controls (left panels) and from three acne vulgaris patients presenting with lesions of increasing severity (right panels) stained with haematoxylin and eosin (H&E), or labelled with antibodies against GATA6, Ki67, and KRT5/6 (brown labelling). Scale bar: 100 μm. b Expression of GATA6 transcripts (210002_at probe) was assessed in unaffected skin and acne lesions from microarray datasets GSE53795 ( n = 12 control skin, n = 12 acne skin; p = 0.00006) and GSE6475 ( n = 12 control skin, n = 6 acne skin; p = 0.112). Mean and individual values are presented. ns not significant; *** p -value < 0.0005; two-tailed unpaired t -test. Source data are provided as a Source Data file.

Article Snippet: Human GATA6 open reading frame (ORF) (NCBI reference sequence NM_005257.5) cloned into a pReceiver-M02 expression plasmid was purchased from GeneCopoeia (Tebu-Bio).

Techniques: Staining, Expressing, Microarray, Control, Two Tailed Test

Journal: Nature Communications

Article Title: Contribution of GATA6 to homeostasis of the human upper pilosebaceous unit and acne pathogenesis

doi: 10.1038/s41467-020-18784-z

Figure Lengend Snippet: a Representative immunofluorescence images of Mock and GATA6-infected IFE keratinocytes labelled with antiGATA6 and phalloidin. Cells were treated with 1 μg ml −1 doxycycline (Dox) for 16 h. b Expression of Ki67 transcripts (average of probes 212020_s_at, 212021_s_at, 212022_s_at, and 212023_s_at) in unaffected skin and acne lesions from microarray datasets GSE53795 ( n = 12 control skin, n = 12 acne skin; p = 0.016) and GSE6475 ( n = 12 control skin, n = 6 acne skin; p = 0.00002). c Proliferation of ORS keratinocytes assessed as % confluence using an Incucyte video-microscope. Values at each time point were normalised to the first scan point of the Mock+Dox condition ( n = 4/condition; p = 0.0001). d Number of colonies (normalised to the number of colonies in the corresponding Vehicle condition) and colony area of ORS keratinocytes overexpressing GATA6 ( n = 18/condition; p < 0.0001 and p = 0.037, respectively). Representative dishes are shown. e Mock-infected or GATA6-infected sebocytes were treated with 1 μg ml −1 Dox for 4 days. Representative histograms of propidium iodide (PI)-stained cells and quantification of % S phase cells compared to the Mock+Dox condition are shown ( n = 13 for Mock conditions, n = 11 for GATA6 + Vehicle, n = 14 for GATA6+Dox; p < 0.000001). f Colony area of Mock or GATA6 SebE6E7 in the presence of 1 μg ml −1 Dox ( n = 6/condition; p = 0.00004). Representative dishes are shown. g Mock or GATA6 infected sebocytes treated with 1 μg ml −1 Dox or vehicle for 4 days ( n = 6 for Vehicle conditions, n = 9 for Dox conditions; p = 0.799). Representative images of TUNEL-stained cells and quantification of the percentage of TUNEL+ cells are shown. Negative (Neg Ctrl, n = 5) and positive (Pos Ctrl, n = 6) controls are shown. a , g Nuclei were counterstained with DAPI. Scale bars: 50 μm. a – g Data are presented as mean ± SD ( c ) or individual values with means ( b , d – g ) and were obtained from two ( a , f ), three ( d , g ), four ( c ), or five ( e ) independent experiments corresponding to n replicates. Statistical analyses were performed with two-tailed unpaired t -test ( b ), linear regression ( c ) or with ordinary one-way ANOVA ( d – g ). ns not significant; * p -value < 0.05; *** p -value < 0.0005; **** p -value < 0.00005. b – g Source data are provided as a Source Data file.

Article Snippet: Human GATA6 open reading frame (ORF) (NCBI reference sequence NM_005257.5) cloned into a pReceiver-M02 expression plasmid was purchased from GeneCopoeia (Tebu-Bio).

Techniques: Immunofluorescence, Infection, Expressing, Microarray, Control, Microscopy, Staining, TUNEL Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Contribution of GATA6 to homeostasis of the human upper pilosebaceous unit and acne pathogenesis

doi: 10.1038/s41467-020-18784-z

Figure Lengend Snippet: a Mock or GATA6-infected keratinocytes treated with 1 μg ml −1 Dox for 5 days and immunolabelled for IVL and LOR ( n = 8/condition; p = 0.011 and p = 0.021 respectively). Representative images and quantitation are shown. b Organ-cultured human HF transfected with siSCR or siGATA6 and maintained for 5 days. Representative images for GATA6 and IVL staining. c – f Mock or GATA6-infected SebE6E7 sebocytes treated with 1 μg ml −1 Dox or vehicle for 4 ( c , e ) or 5 ( d ) days; 1 μg ml −1 Dox and vehicle (DMSO) or 1 μM TRO + 0.1 μM LG for 4 days ( f ). c , e Gene expression in GATA6+Dox sebocytes is represented as log 2 FC versus Mock+Dox sebocytes ( n = 6/condition in c , and n = 20/condition in e ). d Representative images of PLET1 and KRT7 labelling and quantification are shown ( n = 10 for Vehicle conditions and n = 11 for Dox conditions; p = 0.0001 and p = 0.0002, respectively). f Representative images of LipidTOX staining and quantification are shown ( n = 9 for Mock+Dox+Vehicle and n = 10 for other conditions). a – f Scale bars: 50 μm ( f ), 100 μm ( a , b , d ). Nuclei were counterstained with DAPI ( a , b , d , f ). Data are represented as mean ( c , e ) or mean with individual values ( a , d , f ) and were obtained from two ( b , c ), three ( a , d , f ), or five ( e ) independent experiments corresponding to n replicates. Statistical analyses were performed with ordinary one-way ANOVA ( a , c , d , f ) or two-tailed multiple t -tests ( e ). ns not significant; * p -value < 0.05; ** p -value < 0.005; *** p -value < 0.0005; **** p -value < 0.00005. a , c – f Source data are provided as a Source Data file.

Article Snippet: Human GATA6 open reading frame (ORF) (NCBI reference sequence NM_005257.5) cloned into a pReceiver-M02 expression plasmid was purchased from GeneCopoeia (Tebu-Bio).

Techniques: Infection, Quantitation Assay, Cell Culture, Transfection, Staining, Gene Expression, Two Tailed Test

Journal: Nature Communications

Article Title: Contribution of GATA6 to homeostasis of the human upper pilosebaceous unit and acne pathogenesis

doi: 10.1038/s41467-020-18784-z

Figure Lengend Snippet: a Area of colonies formed by SebE6E7 sebocytes cultured with vehicle or 100 nM 5αDHT, 0.1 μM RA, 1 μM RA or 5 μM RA ( n = 10 for Vehicle, n = 6 for 5αDHT, n = 8 for RA conditions). Representative dishes and quantitation are shown. b Sebocytes treated for 5 days with vehicle ( n = 26) or 100 nM 5αDHT ( n = 8), 1 μM RA or 5 μM RA ( n = 12 for RA conditions for GATA6 analysis and n = 10 for RA conditions for LipidTOX analysis). Representative images of LipidTOX and GATA6 labelling and quantitation are shown. Values are expressed as FC versus vehicle condition. c Inhibition of GATA6 expression by shGATA6 lentiviruses. Sebocytes were infected with shSCR or shGATA6 lentiviruses and treated with 1 μg ml −1 Dox with vehicle or 5 μM RA for 5 days ( n = 8 for vehicle conditions, n = 6 for RA conditions). RT-qPCR of GATA6 expression normalised to housekeeping genes. Values are expressed as FC versus shSCR+Dox+Vehicle values. d – f Inhibition of GATA6 by shGATA6 reduces the effects of RA on sebocytes. Cells were treated as in c . d Expression of EMA , PLET1 , KRT17 , and cMYC ( n = 8/condition). e Quantitation of representative images of LipidTOX and GATA6 labelling ( n = 34 for shSCR conditions, n = 14 for shGATA6+Dox+Vehicle, n = 16 for shGATA6+Dox+RA). Values are expressed as FC over shSCR+Dox+Vehicle condition. f Number of colonies and colony area ( n = 10/condition). Representative dishes are shown. b , e Nuclei were counterstained with DAPI. Scale bars: 50 μm ( b , e ). a – f Data are means with individual values and were obtained from three ( a , b , e ) or four ( c , d , f ) independent experiments corresponding to n replicates. Statistical analysis was performed with ordinary one-way ANOVA ( a – f ). ns not significant; * p -value < 0.05; ** p -value < 0.005; *** p -value < 0.0005; **** p -value < 0.00005. a – f Source data are provided as a Source Data file.

Article Snippet: Human GATA6 open reading frame (ORF) (NCBI reference sequence NM_005257.5) cloned into a pReceiver-M02 expression plasmid was purchased from GeneCopoeia (Tebu-Bio).

Techniques: Cell Culture, Quantitation Assay, Inhibition, Expressing, Infection, Quantitative RT-PCR

Journal: Nature Communications

Article Title: Contribution of GATA6 to homeostasis of the human upper pilosebaceous unit and acne pathogenesis

doi: 10.1038/s41467-020-18784-z

Figure Lengend Snippet: a SebE6E7 cells were infected with shGATA6 or shSCR (column 1, n = 12/condition), or with GATA6 or Mock (columns 2–4) lentiviruses and treated with 1 μg ml −1 Dox or vehicle for 3–4 days ( n = 32/condition for column 2, n = 16/condition for column 3, n = 48/condition for column 4). Sebocytes were co-treated with 1 μg ml −1 PGN for 3 days (column 3) or P. acnes for 16 h (column 4). Expression of cytokines and AMP was assessed by RT-qPCR and normalised to housekeeping gene expression. Values are expressed as log 2 FC versus corresponding Mock or shSCR values. b Cytokine protein expression was assessed using cytokine arrays in Mock or GATA6-infected sebocytes treated with Vehicle+Dox or Dox+PGN ( n = 4/condition). Representative arrays are shown. Values are expressed as log 2 FC versus Mock+Dox+Vehicle values for GATA6+Dox+Vehicle condition, or versus Mock+Dox+PGN for GATA6+Dox+PGN values. c Mock and GATA6-overexpressing sebocytes were treated with 1 μg ml −1 Dox and 1 U ml −1 human recombinant IFNγ or vehicle for 3 days. Median intensity of PD-L1 and HLA-DR staining was analysed by flow cytometry ( n = 9/condition). Representative FACS plots are shown. a – c Data are mean ( a ) or mean ± SD ( b ) or mean with individual values ( c ) and were obtained from two ( b ) or at least four ( a , c ) independent experiments corresponding to n replicates. Statistical analyses were performed with two-tailed multiple t -tests ( a ) or ordinary one-way ANOVA ( b , c ). ns not significant; * p -value < 0.05; ** p -value < 0.005; *** p -value < 0.0005; **** p -value < 0.00005. a – c Source data are provided as a Source Data file.

Article Snippet: Human GATA6 open reading frame (ORF) (NCBI reference sequence NM_005257.5) cloned into a pReceiver-M02 expression plasmid was purchased from GeneCopoeia (Tebu-Bio).

Techniques: Infection, Expressing, Quantitative RT-PCR, Gene Expression, Recombinant, Staining, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Contribution of GATA6 to homeostasis of the human upper pilosebaceous unit and acne pathogenesis

doi: 10.1038/s41467-020-18784-z

Figure Lengend Snippet: a , b Bright-field micrographs of SebE6E7 organoids. Sebaceous organoids were labelled for PLET1 and FASN in a . Organoids in b were treated with vehicle (DMSO), 5 μM RA or 25 μM RepSox for 6 days ( n = 110 for Vehicle condition, n = 98 for RA and RepSox conditions). Representative images and quantification of organoid diameter are shown. c Sebaceous organoids were grown as in b and analysed by RT-qPCR. Values were normalised to housekeeping genes and presented as log 2 FC versus vehicle condition ( n = 24 for RepSox condition, n = 26 for RA condition). d Sebaceous organoids formed by mock-, GATA6-expressing, shSCR-expressing or shGATA6-expressing SebE6E7 sebocytes were grown for 7 days in the presence of 1 μg ml −1 Dox. Analysis was performed as in c and values are represented as log 2 FC versus Mock or shSCR conditions ( n = 10/condition). e Sebaceous organoids were grown as in d in the presence of 25 μM RepSox. Analysis was performed as in d and values are represented as log 2 FC versus Mock+Dox+RepSox or shSCR+Dox+RepSox conditions ( n = 10/condition). f , g Cells were treated as in d , e . Representative images for GATA6 and PLET1 ( f ) or IVL and KRT7 ( g ) staining. h GATA6, shGATA6, or control sebaceous organoids were grown for 7 days in the presence of 1 μg ml −1 Dox and stained for P-SMAD2. i Back skin of 60-year-old male labelled for PPARγ and P-SMAD2/3. j KEGG signalling pathways analysis for GATA6 upstream regulators displayed in Supplementary Fig. . a – i Scale bars: 10 μm ( f – h ), 25 μm ( a , b ) and 200 μm ( i ). Nuclei were counterstained with DAPI ( a , f – i ). Data are mean ( c – e ), or mean with individual values ( b ), and were obtained from two ( a , f – h ), three ( d , e ) or five ( b , c ) independent experiments corresponding to n replicates. Statistical analyses were performed with ordinary one-way ANOVA ( b – e ). ns not significant; * p -value < 0.05; ** p -value < 0.005; *** p -value < 0.0005; **** p -value < 0.00005. b – e Source data are provided as a Source Data file.

Article Snippet: Human GATA6 open reading frame (ORF) (NCBI reference sequence NM_005257.5) cloned into a pReceiver-M02 expression plasmid was purchased from GeneCopoeia (Tebu-Bio).

Techniques: Quantitative RT-PCR, Expressing, Staining, Control

Journal: Nature Communications

Article Title: Contribution of GATA6 to homeostasis of the human upper pilosebaceous unit and acne pathogenesis

doi: 10.1038/s41467-020-18784-z

Figure Lengend Snippet: GATA6 marks a population of cells of the human upper pilosebaceous unit that contribute both to the follicular (lower INF/JZ/SD) and sebaceous (upper SG) compartments. GATA6 limits the proliferation and the extent of differentiation within these two compartments. In addition, it negatively regulates IFE/upper INF fate through TGFβ signalling induction. During comedogenesis, GATA6+ progenitors are targeted and GATA6 expression is reduced. This causes a switch in lineage determination within the upper pilosebaceous unit that favours the IFE/upper INF fate.

Article Snippet: Human GATA6 open reading frame (ORF) (NCBI reference sequence NM_005257.5) cloned into a pReceiver-M02 expression plasmid was purchased from GeneCopoeia (Tebu-Bio).

Techniques: Expressing

Journal: Frontiers in Physiology

Article Title: The Allelic Variant A391T of Metal Ion Transporter ZIP8 (SLC39A8) Leads to Hypotension and Enhanced Insulin Resistance

doi: 10.3389/fphys.2022.912277

Figure Lengend Snippet: Murine ZIP8 homology model, metal ion transport and plasma membrane expression of human ZIP8 WT and A391T mutant in transiently transfected HEK293T cells. (A) Left panel: 3D structure of mouse ZIP8 based on the bbZIP X-ray structure . Right panel: Diagram indicating the disposition of C and N-terms and the colouring and numbering of the 8 Transmembrane Helix (TMH). (B) Left panel: Representative fluorescence microscopy images of intracellular Zn 2+ (10 μM) accumulation. Right panel: Normalized values from six independent experiments ( n = 22–51) are represented individually. (C) Left panel: Representative experiment showing the change on fluorescence intensity as result of intracellular Cd 2+ (10 μM) accumulation. Right panel: Normalized values from four independent experiments ( n = 2–30) are represented individually. (D) Left panel: Representative experiment showing 55 Fe 2+ transport kinetics [0.1–10 μM]. Middle panel: Normalized values of iron transport (10 μM) obtained from five independent experiments ( n = 27–40) are represented individually. Right panel: Iron transport (1 μM) in the presence of an excess of different divalent metals (Zn 2+ , Cd 2+ , Co 2+ , Cu 2+ , Mn 2+ and Ba 2+ ) (10 μM). Normalized values from independent experiments ( n = 2) are represented individually. (E) Representative blot showing the plasma membrane surface protein expression determined using an anti-HA monoclonal antibody. Normalized results obtained from three independent experiments, performed in duplicate. (F) Left panel: Biotin content of each sample as loading control. Right panel: Plasma membranes expression of Na + /H + exchanger (NHE-1) and lack of β-actin expression are shown as control of membrane surface samples purity. All the data are mean ± SEM of the indicated number of biological replicates. Statistical differences between groups were assessed using either t -test or Mann-Whitney U according to the sample distribution. Significance was set at p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. NT, non-transfected; EV, empty vector.

Article Snippet: The human SLC39A8 (ZIP8) ORF (NCBI Reference Sequence: NM_022154.5) containing a C-terminal TurboGFP tag, was cloned into the pCMV6-AC-GFP vector (OriGene #RG204200).

Techniques: Expressing, Mutagenesis, Transfection, Fluorescence, Microscopy, MANN-WHITNEY, Plasmid Preparation

Journal: Frontiers in Physiology

Article Title: The Allelic Variant A391T of Metal Ion Transporter ZIP8 (SLC39A8) Leads to Hypotension and Enhanced Insulin Resistance

doi: 10.3389/fphys.2022.912277

Figure Lengend Snippet: ZIP8KI mice express less ZIP8 protein. (A) Sequencing analysis for A393T mutation in the Slc38A9 mice. The 1177G>A mutation in exon 8 results in an exchange of alanine (Ala) to threonine (Thr, blue). For detection of the mutated allele in the genome the SspI restriction site is deleted by a silent exchange of 1185T>C. (B) Genotyping for the detection of the A393T mutation in the Slc39A8 locus. DNA extracted from ear biopsies of mice. The expected PCR signal of 503 bp is cut into two fragments of 371 and 132 bp in the wild type, and the mutant allele remain uncut fragment of 503 bp. The homozygote mouse shows the expected cleavage products, demonstrating introduction of the mutation into the genome. (C) Relative mRNA expression of slc39a8 in lung and (D) kidney of WT and ZIP8KI male mice normalized to actin under StD diet ( n = 8). (E) Western blot analysis and quantification of ZIP8 protein expression in lung and (F) kidney membrane extracts in WT and ZIP8KI ( n = 5–15) Means ± SEM of animals in each experimental condition. ∗ p < 0.05 comparisons made between WT and ZIP8KI mice.

Article Snippet: The human SLC39A8 (ZIP8) ORF (NCBI Reference Sequence: NM_022154.5) containing a C-terminal TurboGFP tag, was cloned into the pCMV6-AC-GFP vector (OriGene #RG204200).

Techniques: Sequencing, Mutagenesis, Expressing, Western Blot

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) Domain diagram of human BLTP2 and of the internally tagged (Halo, EGFP or 2xV5 epitopes) BLTP2. TM indicates the transmembrane region of BLTP2. (B) Schematic model of the arrangement of BLTP2 at contacts of the ER with other membranes. The arrow indicates the site where tags (Halo, EGFP or 2xV5 epitopes) were inserted. Structures are predicted using AlphaFold3. (C and D) U2OS cells (C) and MDA-MB-231 cells (D) stably expressing BLTP2^Halo show enrichment of this protein at ER-PM contacts near the edge of the cell. An optical section close to the basal surface (see dashed red line in the cartoon) is shown. (E) HeLaM cells stably expressing BLTP2^Halo show enrichment of the protein at tubular structures. (F) Localization of endogenous BLTP2 in gene edited HeLaM cells where the 2xV5 epitope was inserted in the coding sequence of BLTP2. Anti-V5 immunofluorescence reveals enrichment of BLTP2 on tubular structures. (G) Validation of the endogenous tagging of BLTP2 by western blotting. Anti-BLTP2 immuno-precipitation coenriched a V5 immunoreactive band. (H) Endogenous BLTP2 (V5 immunoreactivity) co-localizes with exogenous BLTP2^Halo on the same tubular structures. (I) Correlative Light-Electron Microscopy revealed that a BLTP2-positive tubular structure represents a tubular membrane surrounded by ER. (J) BLTP2^Halo localizes at the distal portion of tubular structures that are positive for the PM marker GFP-CAAX and are labeled by the membrane impermeant PM dye CellBrite. (K) TMEM24-mCherry, another ER-PM contact protein, is also present on BLTP2-positive tubular structures but only partially colocalizes with BLTP2^Halo. (L) GFP-MAPPER, an artificial ER-PM tethering protein, is also present on BLTP2-positive tubular structures but does not overlap with BLTP2^Halo. (M) Schematic drawing of BLTP2 localization at contacts of the ER with both the “outer” PM and the distal portions of PM-connected tubular structures.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Stable Transfection, Expressing, Sequencing, Immunofluorescence, Biomarker Discovery, Western Blot, Immunoprecipitation, Electron Microscopy, Membrane, Marker, Labeling

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A and B) U2OS (A) or MDA-MB-231 (B) cells stably expressing BLTP2^Halo show its localization throughout the ER (labeled with ER marker Sec61β) with an enrichment at ER-PM contacts. (C) In some MDA-MB-231 cells, BLTP2^Halo show an enrichment on tubular structures similar to those observed in HeLaM cells. (D and E) BLTP2^Halo mainly localizes throughout the ER in COS-7, as shown by colocalization with the ER marker RFP-Sec61β (D), and only occasionally it is enriched at some tubular structures (E).

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Stable Transfection, Expressing, Labeling, Marker

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) BLTP2-positive tubules are positive for Rab8A. (B) A fluorescent labeled MHC-I antibody added to HeLaM cells expression BLTP^EGFP labels the tubules.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Labeling, Expressing

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) Endogenous BLTP2 localize at the tip of mCherry-Rab10-positive tubular endosomes in HeLaM cells. (B) BLTP2^Halo- and mCherry-Rab10-positive tubular endosomes in HeLaM cells are connected with the PM as revealed by labeling with the membrane impermeant PM dye CellBrite. (C) mCherry-Rab10-positive tubular endosomes in HeLaM cells are also labeled with the PM marker GFP-CAAX. (D and E) Absence of GFP-CAAX positive tubules in HeLaM cells expressing dominant negative Rab10 (mCh-Rab10 T23N). Fluorescence image in (D) and quantification in (E). Two tailed t-test. Mean ±SEM. n=3 independent experiments. 48 cells for WT and 57 cells for the T23N Mutant. (F and G) Expression of dominant negative Rab10 (mCh-Rab10 T23N) abolished BLTP2^EGFP-positive tubules. Fluorescence images in (F) and quantification in (G). Two tailed t-test. Mean ±SEM. n=3 independent experiments. 127 cells for WT Rab10 and121 cells for the T23N mutant. (H) Tubular endosomes positive for endogenous Rab10 immunoreactivity disappear after nocodazole treatment for two hours but are restored after washing out the drug. n=3 independent experiments. Pretreatment: 269 Rab10 tubules from 28 cells; treated: zero tubules observed in the 46 cells examined; washout: 353 Rab10 tubules from 26 cells. (I) Live-imaging of a HeLaM cells after nocodazole washout shows the recovery of a Rab10-positive tubular endosome and the recruitment of BLTP2^EGFP after the tubule reaches the PM and fuses with it. (J) Schematic drawing of BLTP2 recruitment and localization at a Rab10-positive tubular endosome that is continuous with the PM.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Labeling, Membrane, Marker, Expressing, Dominant Negative Mutation, Fluorescence, Two Tailed Test, Mutagenesis, Imaging

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) BLTP2^Halo is enriched at tubular structures positive for the PM marker GFP-CAAX and for an extracellular membrane impermeable dye CellBrite labeled at 37 ℃. (B) The tubular structures of HeLaM cells are also positive for another PM marker, Lyn11-RFP. (C) BLTP2-positive tubules are also labeled by CellBrite at 4 ℃.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Marker, Membrane, Labeling

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) Sequence alignment of the C-terminal region of human BLTP2 with the corresponding region of other species. Conserved residues are highlighted in blue, with darker color indicating higher conservation. a.a.2219-2300 of Drosophila BLTP2 (hobbit) are framed by red solid lines and a.a.2177-2235 of human BLTP2 by magenta dashed lines. (B and C) U2OS cells (B) and HeLaM cells (C) co-expressing the BLTP2 C-terminal deletion mutant BLTP2^EGFP(△59) and BLTP2^Halo full length (FL). BLTP2^EGFP(△59) show reduced localization relative to BLTP2^Halo FL at ER-PM contact sites in U2OS cells (arrowhead) and at tubular structures in HeLaM cells.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Sequencing, Expressing, Mutagenesis

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) PM-connected tubular endosomes in HeLaM cells are positive for PI4P (labeled by iRFP-P4M) and PI(4,5)P 2 (labeled by GFP-PH PLCδ1 ) markers. (B) Design of the rapamycin-dependent dimerization assay to recruit the 4-phosphatase domain of Sac1 (target PI4P) or the 5-phopshatase domain of INPP5E (target PI(4,5)P 2 ) to PM-connected tubular endosomes. (C) BLTP2^Halo disassociates from tubular endosomes after PI4P depletion on their membranes following the recruitment of RFP-FKBP-Sac1. In contrast, BLTP2^Halo show no clear change after PI(4,5)P 2 depletion following the recruitment of RFP-FKBP-INPP5E. (D) BLTP2^Halo gradually disassociates from tubular endosomes after PI4KIIIα inhibition in response to addition of the A1 compound. (E) mCh-Rab10 tubular endosomes persist after BLTP2^Halo disassociation from them in response to A1 treatment. (F) Endogenous BLTP2 also undergoes disassociation from tubular endosomes in response to treatment with A1. (G) Cells in (F) are quantified for the presence of BLTP2 tubules. Two-tailed t-test. Mean ±SEM. n=3 independent experiments. Non treated (NC): 85 cells; A1 treated: 118 cells. (H) BLTP2^Halo re-associates with tubular endosomes after A1 washout.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Labeling, Inhibition, Two Tailed Test

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) The disassociation of BLTP2 after A1 treatment does not correlate with a disruption of the entire Rab10-positive tubular endosome network in HeLaM cells. (B) BLTP2 re-establish contact with tubular endosomes after A1 washout.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Disruption

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A and B) Interaction diagram exported from the Yeast Interactome Website ( http://yeast-interactome.biochem.mpg.de:3838/interactome/ ) revealing protein interactions of the BLTP2 yeast orthologues Fmp27 (A, score=10) and Ypr117w (B, score=3) with Ybl086c, the ortholog of mammalian FAM102A/B. (C) Domain organization of Ybl086c with FAM102A/B comprising an N-terminal C2 domain and a C-terminal disordered region. The AlphaFold3-predicted BLTP binding site in FAM102A and B is shown in (J) (see also ). (D) Solo expression of FAM102A-GFP shows localization at the PM in U2OS cells, with no focal accumulations as expected for ER-PM contact sites. (E) In U2OS cells co-expressing FAM102A-GFP and BLTP2^Halo, the two proteins colocalize at ER-PM contacts. (F) In COS-7 cells, where BLTP2 does not accumulate at ER-PM contacts when expressed alone, co-expression of FAM102A-GFP recruits BLTP2^Halo to ER-PM contacts. (G) Solo expression of FAM102B-GFP in U2OS cells results in its diffuse localization. (H) In U2OS cells co-expressing FAM102B-GFP and BLTP2^Halo, the two proteins colocalize at ER-PM contacts. (I) In COS-7 cells, where BLTP2 does not accumulate at ER-PM contacts when expressed alone, co-expression of FAM102B-GFP recruits BLTP2^Halo to ER-PM contacts. (J) AlphaFold3 predicts an interaction between the C-terminal region of FAM102A (magenta) and a two-helices hairpin (green) (Fam102 Binding Motif, FBM) projecting out of the BLTP2 rod-like core (blue). (K and L) COS-7 cells co-expressing BLTP2^EGFP and BLTP2^Halo-△FBM with FAM102A-iRFP (K) or FAM102B-iRFP (L), respectively, showing that BLTP2^Halo-△FBM is not co-enriched with WT BLTP2^EGFP at ER-PM contact sites, but remains diffuse throughout te ER. (M and N) In COS-7 cells co-expressing BLTP2^Halo, the M1R receptor and either FAM102A-GFP (M) or FAM102B-GFP (N), BLTP2^Halo disassociates from the ER-PM contact sites and redistributes to the entire ER in response to PI(4,5)P 2 depletion by Oxo-M addition. BLTP2^Halo re-localizes to ER-PM contacts after addition of the Oxo-M antagonist atropine.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Binding Assay, Expressing

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) Sequence alignment of human FAM102A and FAM102B where their N-terminal C2 domains (purple dashed box) and their C-terminal predicted BLTP2 interacting regions (red dashed box) are highlighted. (B) Co-localization of FAM102A-GFP (Top) and FAM102B-GFP (Bottom) with BLTP2^Halo at ER-PM contact sites in MDA-MB-231 cells. (C) AlphaFold3 predicted interaction between FAM102B and BLTP2. (D) FAM102A-GFP recruits BLTP2^Halo to ER-PM contact sites in HeLaM cells.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Sequencing

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) Solo expression of FAM102A-GFP in HeLaM cells results in its diffuse localization, while solo expression of FAM102B-GFP results in its enrichment localization on tubular structures similar to BLTP2 localization. (B and C) FAM102B-GFP co-localizes with either BLTP2^Halo (B), or endogenous BLTP2 (C) on tubular structures in HeLaM cells. (D-F) Co-expression of BLTP2^EGFP FL (full length) with either BLTP2^Halo FL or BLTP2^Halo-△FBM, respectively, in HeLaM cells, showing that deletion of the FBM motif decreases the enrichment of BLTP2 at the tubular structures (D). (E) shows the method used for the quantification. The quantification result is shown in (F). Two-tailed t-test. Mean ±SEM. (FL) n=63 tubules from nine cells, (△FBM) n=111 tubules from 15 cells. (G) GFP-FBP17 localizes on the same BLTP2^Halo-positive tubules but does not overlap with the BLTP2^Halo signal. (H and I) GFP-mAmph2 co-localizes with BLTP2^Halo on the tubular structures (H), while GFP-mAmph2-BAR* (I) localizes on the same tubules but does not overlap with BLTP2 ^Halo. (J) AlphaFold3 predicts the interaction of a loop of BLTP2 (loop1) with the SH3 domain of amphiphysin. This loop harbors a RAPHPP sequence (RxP motif) which fits an SH3 domain binding consensus. (K and L) Co-expression of BLTP2^EGFP FL with a BLTP2^Halo construct in which each of these six amino acids of the SH3 binding consensus were mutated to alanine (BLTP2^Halo RP→AA ), or BLTP2^Halo RP→AA with the additional deletion of the FBM (BLTP2^Halo RP→AA -△FBM), showing a synergistic effect of abolishing SH3 domain binding and FAM102 binding in reducing the targeting of BLTP2 to the tubules. The quantification result is shown in (L). One way ANOVA. Mean ±SEM. (FL) n=63 tubules from nine cells, (RP→AA) n=103 tubules from 13 cells, (RP→AA and △FBM) n=97 tubules from 13 cells. (M) Schematic drawing depicting co localization of BLTP2, FAM102B and amphiphysin 2 on the PM-connected tubular endosomes, but segregation form FBP17.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Expressing, Two Tailed Test, Sequencing, Binding Assay, Construct

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) COS-7 cells showing that BLTP2^Halo is recruited to PM tubular invaginations induced by expression of CFP-mAmph2. (B) COS-7 cells showing that BLTP2^Halo is not recruited to PM tubular invaginations induced by expression of GFP-mAmph2 BAR* which lacks the SH3 domain. (C) COS-7 cells showing that BLTP2^Halo is not recruited to PM tubular invaginations induced by expression of GFP-FBP17. (D) Sequence alignment of the region comprising the RAPHPP motif among BLTP2 from several organisms. The RAPHPP motif is conserved among mammalian species (blue arrowhead).

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Expressing, Sequencing

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A and B) COS-7 cells showing macropinosomes (labeled by internalized fluorescent anti MHC-I antibodies) that undergo a dramatic morphological change and acquire BLTP2^EGFP as they fuse and collapse with the PM. In (B), a newly formed macropinosome at first loses PI(4,5)P 2 , but then reacquires PI(4,5)P 2 as it fuses with the PM. (C) Schematic drawing of macropinosome recycling. A nascent macropinosome first loses PI(4,5)P 2 after fission from the PM, then it acquires the identity of early endosomes (PI3P, APPL and Rab5), or regain PI(4,5)P 2 as it fuses back to the PM. (D) Stimulation of macropinocytosis by expression of HRas G12V in COS-7 cells. BLTP2^Halo is recruited to a newly formed macropinosome that loses PI(4,5)P 2 but does not acquire APPL2 and regains PI(4,5)P 2 signaling during its recycling back to the PM. (E) Schematic drawing depicting the assay to monitor macropinosome fusion with the PM. PM dye is added after the formation of macropinosomes. Pre-formed macropinosomes will not be labeled by the dye until they fuse with the PM. (F) A BLTP2-positive macropinosome gains access to the PM dye showing that it fuses with the PM. Macropinocytosis was stimulated by expressing HRas G12V. (G) Acute recruitment of BLTP2^Halo to a macropinosome in the process of fusing with the PM in HeLaM cells. The macropinosome is also positive for mCherry-Rab10. Scale Bar 1μm. (H) Schematic drawing of the recruitment of BLTP2 to a recycling macropinosome undergoing fusion with the PM.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Labeling, Expressing

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) Acute recruitment of BLTP2^EGFP to macropinosomes (labeled by internalized fluorescent anti MHC-I antibodies) in the process of fusing with the PM. (B) Z-stack of the time lapse images shown in proving that the macropinosome is not moving out from the focal plane during the imaging session.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Labeling, Imaging

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) PH AKT also label nascent macropinosomes. The recycling macropinosomes are positive for PH AKT while recruiting BLTP2 and are not positive for APPL1. (B) BLTP2-positive recycling macropinosomes are not positive for early macropinosome marker Rab5. (C) BLTP2-positive recycling macropinosomes are positive for exocytic factor Rab8A. (D) After treating cells with VPS34 inhibitor SAR405, the PI3P positive early macropinosomes can reverse to PI(4,5)P 2 and recycle back to the PM.

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Marker

Journal: bioRxiv

Article Title: Multiple interactions mediate the localization of BLTP2 at ER-PM contacts to control plasma membrane dynamics

doi: 10.1101/2025.02.07.637094

Figure Lengend Snippet: (A) Western blot validating knock-out (KO) of BLTP2 in HeLaM cells. Endogenouse BLTP2 is enriched using IP before detection. Three independent KO clones are verified. (B) BLTP2-KO HeLaM cells showing presence of PI(4,5)P 2 -positive intracellular vacuoles. Tubular recycling endosomes are still present in these cells. (C and D) Expression of HRas G12V in HeLaM cells (two independent clones: KO-2C3 and KO-2C6) induces formation of macropinosomes/intracellular vacuoles in both WT and BLTP2-KO cells. However, only in the KO cells a large fraction of these vesicles remain PI(4,5)P 2 positive. Quantification of PI(4,5)P 2 positive macropinosomes is shown in (D). One way ANOVA. Mean ±SEM. n=3 independent experiments. 123 cells for WT, 137 cells for KO-2C3, 161 cells for KO-2C6,119 cells for KO-2C3 rescue and 122 cells for KO-2C6 rescue. (E and F) BLTP2-KO cells expressing HRas G12V together with FAM102A-GFP (E), or FAM102B-GFP (F). FAM102A-GFP, but not FAM102B-GFP, is enriched on PH PLCδ1 labeled PI(4,5)P 2 positive macropinosomes. (G and H) PM dye (G), or 10KD Dextran-488 (H) were added to BLTP2-KO cells expressing HRas G12V. A fraction of the pre-existing PI(4,5)P 2 positive vacuoles were labeled by the dye within minutes after PM dye addition (G), or showing internalization of Dextran-488 (H). (I) Schematic model of BLTP2 localization at contacts of the ER with PM and PM-connected structures (left). Illustration of the molecular interaction of BLTP2 with phosphoinositides and its binding proteins (FAM102A/B, N-BAR domain proteins) at these contact sites (right).

Article Snippet: The original clone containing the BLTP2 ORF (NCBI reference sequence: NM_014680.5) was obtained from GenScript.

Techniques: Western Blot, Knock-Out, Clone Assay, Expressing, Labeling, Binding Assay