EMBO/SourceData huggingface dataset:
Figure legend: "Western blots against NPC1 actin-β (ACTB) in total extracts (T) and biotin-streptavidin immunoprecipitates (S) from synaptosomes from wt and NPC1nmf164 mice in which cLTP was induced or not. Graphs show mean ± SEM of the levels of biotinylated-surface NPC1 as a percentage of the total amount of each protein (n=3 mice, 3 month-old, 2-way ANOVA, *pNPC1 = 0.0251"
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "Western blots against CYP46A1 and actin-β (ACTB) in total extracts (T) and biotin-streptavidin immunoprecipitates (S) from synaptosomes from wt and NPC1nmf164 mice in which cLTP was induced or not. Graphs show mean ± SEM of the levels of biotinylated-surface CYP46A1 as a percentage of the total amount of each protein (n=3 mice, 3 month-old, 2-way ANOVA , *pCYP46A1 = 0.0371)."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "F. Representative images of GluA1 immunocytochemical staining before and after cLTP in non-permeabilized (surface) and permeabilized (total) cultured hippocampal neurons expressing sh-scr or sh-NPC1 RNAs. Graph shows mean ± SEM GluA1 surface staining with respect to total in cLTP conditions, as a percentage of the sh-scr baseline (n=20 images per condition from 2 different experiments, 2-way ANOVA, ***psh-scr &lt; 0.001)."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "A. Western blots against CYP46A1 and ACTB in extracts of total brain and synaptosomes from wt and NPC1nmf164 mice as a percentage of wt values. Graphs show 3 biological replicates mean ± SEM of CYP46A1 level normalized to ACTB in arbitrary units."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "B. Quantitative PCR of Cyp46 of total brain and synaptosomes from wt and NPC1nmf164 mice. 3 biological replicates mean ± SEM is shown."
NCBI gene IDs:
"Cyp46: 13116
NPC1: 18145"
Figure legend: "C. Mean ± SEM cholesterol levels in EFV-treated or non-treated synaptosomes from wt (n=5 mice, 14-week-old, unpaired Student t test, *p = 0.0233) and NPC1nmf164 (n=4 mice, 14 week-old, unpaired Student t test, *p = 0.0188) mice. EFV values are expressed as percentage of their corresponding non-treated controls in wt and NPC1nmf164 conditions."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "D. Basal synaptic transmission in EFV-treated and non-treated hippocampal slices from wt and NPC1nmf164 mice expressed as mean ± SEM EPSP slope (n=9 slices from 5 WT mice, n=11 slices EFV-treated from 5 WT mice, n=8 slices from 5 NPC1nmf164 mice and n=13 slices EFV-treated from 5 NPC1nmf164 mice, all mice 14-week-old, 2-way ANOVA, *p = 0.0117)."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "E. LTP in EFV-treated and non-treated hippocampal slices from wt and NPC1nmf164 mice expressed as mean ± SEM percentage of EPSP slope over baseline (n=7 slices from 5 WT mice, n=9 slices EFV-treated from 5 WT mice, n=10 slices from 5 NPC1nmf164 mice and n=11 slices EFV-tretaed from 5 NPC1nmf164 mice, all mice 14-week-old, 2-way ANOVA, ***p &lt; 0.0001)."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "F. Mean ± SEM paired pulse facilitation in EFV-treated and non-treated hippocampal slices from wt and NPC1nmf164 mice (n=11 slices from 5 WT mice, n=9 slices EFV-treated from 5 WT mice, n=10 slices from 5 NPC1nmf164 mice and n=11 slices EFV-tretaed from 5 NPC1nmf164 mice, all mice 14-week-old, 2-way ANOVA, **pNPC1nmf164=0.0027, *pNPC1nmf164+EFV=0.0229)."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "Representative images of GluA1 immunocytochemical staining before and after cLTP induction in non-permeabilized (surface) and permeabilized (total) cultured hippocampal neurons expressing sh-scr or sh-NPC1 RNAs and treated or not with EFV. Graph shows the mean ± SEM GluA1 surface staining with respect to total after cLTP, as a percentage of the sh-scr baseline (n=20 images per condition from 2 separate experiments, 2-way ANOVA, ***psh-scr &lt; 0.001, **psh-scr+EFV &lt; 0.01, ***psh-NPC1+EFV &lt; 0.001)."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "The following analyses were assessed in wt and NPC1nmf164 mice treated or not with 0.09mg/kg/day EFV A. Mean ± SEM 24(S)-hydroxycholesterol plasma level after 6 and 8 weeks of oral EFV treatment in wt (n=6 mice, 14 week-old and 16 week-old, 2-way ANOVA, **p6 weeks = 0.0023) and NPC1nmf164 (n=5 mice, 14 week-old and 16 week-old, 2-way ANOVA, ***p6 weeks &lt; 0.0001; ***p8 weeks &lt; 0.0001) mice."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "The following analyses were assessed in wt and NPC1nmf164 mice treated or not with 0.09mg/kg/day EFV B. Mean ± SEM body weight in grams in wt and NPC1nmf164 mice treated or not with EFV (n=4 mice, unpaired Student t-test, ***pNPC1nmf164 &lt; 0.0001, **pNPC1nmf164+EFV = 0.0048)"
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "The following behavioural tests were assessed in wt and NPC1nmf164 mice treated or not with 0.09mg/kg/day EFV C. Mean ± SEM discrimination index as a normalized ratio of time spent with novel and familiar objects in the Object Placement Recognition test (n=6 mice, 10 week-old, 2-way ANOVA, **pWT/NPC1nmf164 = 0.0065, ***pNPC1nmf164/NPC1nmf164+EFV = 0.0008)."
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "The following behavioural tests were assessed in wt and NPC1nmf164 mice treated or not with 0.09mg/kg/day EFV D. Mean ± SEM percentage of entries in the novel arm of the Y maze test (n=10 mice, 10 week-old, 2-way ANOVA, ***p &lt; 0.0001). E. Mean ± SEM percentage of freezing time in the Contextual Fear Conditioning test (n=10 mice, 10 week-old, 2-way ANOVA, **pNPC1 &lt; 0.0025, *pNPC1+EFV &lt; 0.0181). F. Mean ± SEM percentage of freezing time in the Cued Fear Conditioning test (n=10 mice, 10 week-old, 2-way ANOVA, *pNPC1 &lt; 0.0453, **pNPC1+EFV &lt; 0.0054). "
NCBI gene IDs:
"NPC1: 18145"
Figure legend: "The following analyses were assessed in wt and NPC1nmf164 mice treated or not with 0.09mg/kg/day EFV G. Mean ± SEM cholesterol level in synaptosomes of wt and NPC1nmf164 mice treated or not with EFV expressed as percentage of the wt non-treated samples (n=4 mice, 14 week-old, 2-way ANOVA, *pNPC1 &lt; 0.0476, *pNPC1+EFV &lt; 0.0118)."
NCBI gene IDs:
"NPC1: 
NPC1: 18145"
Figure legend: "The following analyses were assessed in wt and NPC1nmf164 mice treated or not with 0.09mg/kg/day EFV H. Representative fluorescence images of the CA1 hippocampal region from wt and NPC1nmf164 mice treated or not with EFV and stained with filipin and an antibody against LAMP1. White arrows indicate lysosome enlargement in the hippocampus of non-treated NPC1nmf164 mice. Graph to the right shows mean ± SEM fluorescence intensity associated to filipin per area in arbitrary units (n=4 mice, 14-week old, unpaired Student t-test, **pNPC1nmf164 = 0.0011, *pNPC1nmf164+EFV = 0.0273)). I. Survival graph for wt and NPC1nmf164 mice treated or not with EFV (n=5 mice, 2-way ANOVA, ***p &lt; 0.0001). "
NCBI gene IDs:
"NPC1: 
NPC1: 18145"
Figure legend: "(a) Cells were transfected with control siRNA or with siRNA specific to IRGM (top) and DRP1 (bottom) for 48 h and protein samples from these cells were then analysed by immunoblotting with anti-IRGM and anti-DRP1 antibodies. Actin was used as a loading control."
NCBI gene IDs:
"DRP1: 10059
IRGM: 345611"
Figure legend: "(b) HeLa cells were treated with siRNA specific to either IRGM or DRP1 as indicated and were then labelled with MTR and analysed by live-cell confocal microscopy. Scale bars, 5 μm."
NCBI gene IDs:
"DRP1: 10059
IRGM: 345611"
Figure legend: "(c) Cells were treated with either control siRNA or with DRP1, IRGM, ATG7 or BECN1 (Beclin 1) siRNAs, labelled with MTR and the percentage of cells with mitochondrial morphologies ranging from normal, punctiform (dots) and elongated were quantified. Definition of mitochondrial morphologies and quantification criteria are given in Supplementary Information, Figure S2."
NCBI gene IDs:
"ATG7: 10533
BECN1: 8678
DRP1: 10059
IRGM: 345611"
Figure legend: "(d) Cells were transfected with either control siRNA or with siRNA specific to MFN1/MFN2, IRGM or IRGM and MFN1/MFN2, and then labelled with MTR and analysed for mitochondrial morphologies. Data are means ± s.e.m. (c, n = 3; d, n = 4). Asterisk indicates P 0.05, double asterisks indicate P 0.01 and dagger indicates P &gt; 0.05 (t-test)."
NCBI gene IDs:
"IRGM: 345611
MFN1: 55669
MFN2: 9927"
Figure legend: "(a) U937 cells were transfected with siRNA specific to FIS1, DRP1 or IRGM (or control oligonucleotides) as indicated, followed by plasmid encoding GFP-LC3. Autophagy was induced by treatment with hIFN-γ. GFP-LC3 puncta in cells were imaged by confocal microscopy. Scale bars, 5 μm. (b) LC3 puncta per cell were quantified from cells treated as in a."
NCBI gene IDs:
"DRP1: 10059
FIS1: 51024
IRGM: 345611
LC3: 84557"
Figure legend: "(c) U937 cells were transfected with siRNA specific to FIS1, DRP1 or IRGM (or control oligonucleotides) as indicated, followed by plasmid encoding GFP-LC3. Autophagy was induced by starvation. GFP-LC3 puncta in cells were imaged by confocal microscopy. Scale bars, 5 μm."
NCBI gene IDs:
"DRP1: 10059
FIS1: 51024
IRGM: 345611
LC3: 84557"
Figure legend: "(d) U937 cells were transfected with either control siRNA or siRNA specific to MFN1/MFN2, endogenous LC3 puncta per cell were quantified."
NCBI gene IDs:
"MFN1: 55669
MFN2: 9927"
Figure legend: "(b) HeLa cells were transfected with plasmids encoding fluorescent protein-tagged IRGMb, IRGMd WT (wild-type IRGMd) or IRGMS47N mutant (control cells; vector expressing GFP only), and after 48 h were labelled with MTR and imaged by live-cell confocal microscopy. Scale bars, 5 μm."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(c) Fluorescence intensity analysis of green and red channels along a line drawn through two adjacent cells, one GFP-IRGMd positive and one GFP-IRGMd negative, as indicated in the inset."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(g) Nitrocellulose filters spotted with the indicated lipids (defined in e) were incubated with wild-type IRGMd, IRGMdS47N, and the IRGM isoform IRGMb at the indicated concentrations. GST control is shown in Supplementary Information, Figure S1d. Membranes in e-g were probed with anti-GST antibody."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(a) HeLa cells transfected with plasmids encoding GFP or GFP-IRGMd were stained with MTR and imaged by live-cell confocal microscopy. Scale bars, 5 μm. (b-d) Quantification of mitochondrial morphologies in cells transfected with IRGMd, IRGMdS47N or IRGMb GFP fusions (control cells express GFP only)."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(e)HeLa were transfected with the indicated siRNAs and vector encoding GFP-IRGMd, and labelled with MTR. The percentage of cells with mitochondrial morphologies ranging from normal, punctiform (dots) and elongated were quantified."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(f) HeLa were transfected with the indicated siRNAs and vector encoding GFP-IRGMd (or vector encoding GFP only, as a control) and labelled with MTR. Percentages of GFP+ cells with MTR staining at 48 h were quantified."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(g) Confocal microscopy images of HeLa cells transfected with vector encoding GFP-DRP1 (A) or GFP-IRGMd (B-G) and labelled with MTR (A-D) or antibodies against COX-IV (F). A, steady-state DRP1 distribution; B-D, translocation of IRGMd from the cytosol to mitochondria at indicated times post-transfection; E-G, co-localization of IRGMd and COX-IV (G is merged from E and F). Scale bars, 5 μm."
NCBI gene IDs:
"DRP1: 10059
IRGM: 345611"
Figure legend: "(h) HeLa cells, co-transfected with GFP-IRGMd, and control, BECN1 or ATG7 siRNA, were labelled with MTR 48 h post-transfection, imaged by live-cell microscopy, and percentages of GFP+ cells that were also MTR+ were quantified."
NCBI gene IDs:
"ATG7: 10533
BECN1: 8678
IRGM: 345611"
Figure legend: "(i) Cells from Atg7 wild-type (WT) or Atg7−/− MEFs transfected with vectors encoding GFP-IRGMd for 48 h were stained with MTR, imaged, and percentage of GFP+ cells that were also MTR+ cells was quantified."
NCBI gene IDs:
"Atg7: 74244
IRGM: 15944"
Figure legend: "(j) Wild-type W2 (Bax/Bak+/+) or mutant D3 (Bax/Bak−/−) BMK cells were transfected with vectors encoding GFP-IRGMd and after 48 h were labelled with MTR. Cells were imaged by live-cell microscopy, and percentages of GFP+ cells that were also MTR+ were quantified."
NCBI gene IDs:
"Bak: 12018
Bax: 12028
IRGM: 15944"
Figure legend: "(k) HeLa cells were transfected with vectors encoding GFP-IRGMd, treated with z-VAD as indicated, and after 48 h stained with MTR. Cells were imaged by live-cell microscopy, and percentages of GFP+ cells that were also MTR+ were quantified. Data are means ± s.e.m. (n = 3). Dagger indicates P ≥ 0.05, asterisk indicates P 0.05 and double asterisks indicate P 0.01 (t-test) for the indicated data."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(a) Percentage of HeLa cells transfected with vectors encoding GFP or GFP-IRGMd that were rounded 48 h post-transfection."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(b) HeLa cells transfected with plasmids encoding GFP or GFP-IRGMd were stained with 7-AAD after 48 h and percentages of GFP+ cells that were also 7-AAD+ were quantified."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(c) HeLa cells transfected with vectors encoding GFP-IRGMd versus GFP-IRGMdS47N and YFP-IRGMb (vector encoding GFP only was transfected as a control) were stained with 7-AAD after 48 h, and percentages of GFP+ (for IRGMd and IRGMS47N) or YFP+ (for IRGMb) cells that were also 7-AAD+ were quantified."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(d) HeLa cells were transfected with plamsids encoding GFP-IRGMd, GFP-IRGMDS47N or YFP-IRGMb (vector encoding GFP only was transfected as a control), and after 48 h were stained with propidium iodide (PI). Percentages of GFP+ or YFP+ cells that were also PI+ were quantified."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(e) Images of HeLa cells transfected with vectors encoding GFP-IRGMd (bottom) were stained for active caspases 3 and 7 using FLICA dye. Staurosporine (STS; middle images) was used as a positive control. Cells were transfected with vectors encoding GFP only as a control (top)."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(f) HeLa cells were transfected with plasmids encoding GFP-IRGMd and were stained with antibodies against HMGB1 (absence of nuclear HMGB1 stain is a marker of HMGB1 release) or DAPI. Top: images indicate GFP-IRGMd (left), HMGB1 (middle) and DAPI (right) localization. Bottom: phase-contrast microscopy image of cells (left) and image merged from other panels (right). Scale bars, 5 μm."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "(g-i) Cells were transfected with plasmids encoding GFP or GFP-IRGMd, and control or DRP1 siRNA, as indicated. After 48 h, (g) cells were stained with 7-AAD and percentages of GFP+ cells that were also 7-AAD+ were quantified, (h) rounded cells were quantified, or (i) cells were stained with HMGB1 and HMGB1+ nuclei were quantified. Data are means ± s.e.m. (n = 3). Asterisk indicates P 0.05 and double asterisk indicate P 0.01 (t-test)."
NCBI gene IDs:
"DRP1: 10059
IRGM: 345611"
Figure legend: "(a, b) HeLa cells were transfected with plasmids encoding GFP, CFP-IRGMa, GFP-IRGMc or GFP-IRGMd, and after 24 h (a) or 48 h (b), stained with MTR. Percentages of positively transfected cells that were also MTR+ were then quantified. (c, d) Cells were transfected as in a, b, and after 24 h (c) or 48 h (d) the percentages of positively transfected cells that were also rounded by morphology were quantified."
NCBI gene IDs:
"IRGM: 345611"
Figure legend: "A-D. Example expression patterns for select genes identified by applying a general linear model (glm) including state, genotype, and interaction terms. Dots represent individual biological replicates (N = 3); lines highlight changes in mean values for replicates between each state (log2FC &gt;1 and FDR &lt; 0.05). A. State dependent expression is exemplified by Nanog which showed no difference between strains. B. Expression of Zfp277 exemplifies strain-dependence being consistently higher in B6. C &amp; D. A significant genotype x state (GXS) interaction was identified for Nr5a2 (C, a marker for pluripotency) and Sox1 (D, a marker for neuronal differentiation)."
NCBI gene IDs:
"Nanog: 71950
Nr5a2: 26424
Sox1: 20664
Zfp277: 246196"
Figure legend: "G. Coverage profile for Gstp2 locus on Chr 19 showing H3K4me3, ATAC, and RNA read depth from merged replicates. Locations of Chr 7 trans caQTL targets are indicated with black bars and LOD scores listed below."
NCBI gene IDs:
"Gstp2: 14869"
Figure legend: "F. Left - Coverage profiles (averaged biological replicates, N=3) for ChIP factor occupancy at Cd59a on Chr 2 under trans-regulation by the Chr 13d QTL. Distal ca-QTL target region used for quantification is indicated by a black box with LOD score below. Right - Scatterplots for ChIP factors comparing quantitative level of modification/binding for independent replicates between B6 and D2 ESCs."
NCBI gene IDs:
"Cd59a: 12509"
Figure legend: "I. Similar to F, coverage profiles for ChIP factor occupancy for a putative regulatory element on Chr 5 near Zfp932, targeted by Chr 12 trans caQTL, showed greater occupancy of POU5F1 and increased open chromatin compared to D2. Distal ca-QTL target region used for quantification is indicated by a black box with LOD score below."
NCBI gene IDs:
"Zfp932: 69504"
Figure legend: "A. Coverage profile for H3K4me3, ATAC, P300, and TRIM28 ChIP at a target locus for the QTL on Chr 4 for both chromatin accessibility (black boxes) and gene expression (Riok3 and Rmc1). LOD scores are listed under each target locus."
NCBI gene IDs:
"Riok3: 66878
Rmc1: 76482"
Figure legend: "(A-C) Atg5 KO MEFs were either untreated (NT) or starved (ST) with EBSS (Earle's Balanced Salt Solution) for 30 min. Total membranes (mem) from lysed cells were collected and incubated in a lipidation reaction with cytosols prepared from starved HEK293T cells. Reactions contained the indicated concentrations of PI3K inhibitor (PI3KI) 3-methyladenine (3-MA) (B) or FYVE protein (C). A diagram of the experimental scheme is shown in (A). RPN1, Ribophorin 1"
NCBI gene IDs:
"Atg5: 11793"
Figure legend: "(D, E) Atg5 KO MEFs were either untreated (NT) or starved (ST) with EBSS in the absence or presence of 20 nM wortmannin (Wtm) or 10 mM 3-methyladenine (3-MA) for 30 min. Membranes from each treated cell sample were collected and subjected to a differential centrifugation to separate the 3K ×g, 25K ×g and 100K ×g pellet fractions followed by a lipidation assay as above (E). A diagram is shown in (D)."
NCBI gene IDs:
"Atg5: 11793"
Figure legend: "(F, G) Atg5 KO MEFs were starved for 30 min. Membranes in the 25K ×g and 100K ×g pellets from a differential centrifugation were collected as described above. A similar lipidation assay was performed in the presence of indicated concentrations (Conc in G) of 3-MA, wortmannin (F) and FYVE protein as well as a PI3P binding-deficient FYVE mutant protein (C/S) (G). Quantification of lipidation activity is shown as the ratio of LC3-II to LC3-I (II/I)."
NCBI gene IDs:
"Atg5: 11793
FYVE: 57590///53349"
Figure legend: "(B) A small vesicle generation assay as above was performed with the indicated conditions. Slowly-sedimenting vesicles were collected to determine activity in the LC3 lipidation reaction. 3-MA, 5 mM; Wortmannin, 20 nM; S, starved Atg5 KO MEF cytosol; N, untreated Atg5 KO MEF cytosol; −/AP, in the absence of ATP regeneration and GTP and in the presence of apyrase (AP)."
NCBI gene IDs:
"Atg5: 11793"
Figure legend: "(F) A small vesicle generation assay as shown in (A) was performed with cytosols from starved Atg5 KO MEF from control or Sec23A knockdown cells in the absence or presence of Sar1A (H79G) or 3-MA followed by a lipidation reaction."
NCBI gene IDs:
"Atg5: 11793///11793
Sec23A: 20334"
Figure legend: "(G) Atg5 KO MEFs were starved for 30 min. Differential centrifugation as shown in Figure 1D was performed to collect the 25K ×g and 100K ×g pellet fractions. The LC3 lipidation was performed on these two fractions with indicated concentrations of SAR1 (H79G). Quantification of lipidation activity is shown as the ratio of LC3-II to LC3-I (II/I)."
NCBI gene IDs:
"Atg5: 11793"
Figure legend: "(A) Atg5 KO MEFs were either untreated or starved with or without 20 nM wortmannin for 30 min. Cells were harvested and ERGIC membranes were isolated by pooling fractions 3 and 4 (F2 in Figure 2C,D) of the OptiPrep gradient in the three-step fractionation approach followed by immunoblot to examine the amount of indicated markers on ERGIC membranes."
NCBI gene IDs:
"Atg5: 11793"
Figure legend: "(B) Atg5 KO MEFs were transfected with a plasmid encoding an HA-tagged SEC31A. After transfection (24 hr), the cells were treated as shown in (A). Immunofluorescence was performed with anti-ERGIC53 and anti-HA antibodies and the cells were examined by confocal microscopy. Bar: 10 μM (C) Quantification of SEC31A overlapped with ERGIC53 shown in (B) using the Pearson's correlation coefficient (PCC). Error bars represent standard deviations from ∼20 cells in three independent experiments. p values were calculated by T-test."
NCBI gene IDs:
"Atg5: 11793"
Figure legend: "A The PA synthesis defective mutants pldα1 pldδ-1 and pldα1 pldδ-2 are salt-sensitive. Six-d-old seedlings were grown in half-strength Murashige and Skoog (MS), and then transferred to half-strength MS medium with or without 100 mM NaCl for 7 d."
NCBI gene IDs:
"pldα1: 820816
pldδ: 829733"
Figure legend: "G, H Analysis of the average values of the net Na+ (G) and K+ (H) fluxes of Col-0, sos2, C-1 and C-2. Six-d-old seedlings were treated with or without 50 mM NaCl for 24 h. Error bars represent SD (n ≥ 3)."
NCBI gene IDs:
"sos2: 833502"
Figure legend: "I-N The contents of shoot Na+ (I), shoot K+ (J), root Na+ (K) and root K+ (L) as well as the relative ratios of shoot Na+/K+ (M) and root Na+/K+ (N) of Col-0 and pldα1 pldδ mutants. Six-d-old seedlings were transferred to half-strength MS medium with or without 100 mM NaCl treatment for 7 d. Error bars represent SEM of three independent biological repeats. O-T The contents of shoot Na+ (O), shoot K+ (P), root Na+ (Q) and root K+ (R) as well as the relative ratios of shoot Na+/K+ (S) and root Na+/K+ (T) of Col-0, sos2, C-1 and C-2 mutants. Six-d-old seedlings were transferred to half-strength MS medium with or without 50 mM NaCl treatment for 7 d. Error bars represent SEM of three independent biological repeats. D"
NCBI gene IDs:
"pldα1: 820816
pldδ: 829733
sos2: 833502"
Figure legend: "C PA promotes the activity of SOS2 under salt stress. Ten-d-old transgenic plants expressing 35S:6×Myc-SOS2 in Col-0 or pldα1 pldδ-1 background were treated with or without 100 mM NaCl or 50 μM PA for 12 h. The total protein was extracted and immunoprecipitated with anti-Myc antibody-conjugated agarose, and then the purified Myc-SOS2 proteins at similar protein levels were incubated with His-SCaBP8 in the kinase assays. His-SCaBP8 was used as a specific substrate of SOS2. The numbers below the blots indicate the relative ratios of the signal intensity between the autoradiograph and Myc-SOS2 bands (Autorad/Myc). Statistical analysis of the relative ratios is shown D PA promotes the PM localization of SOS2 under salt stress. Ten-d-old seedlings were treated as described in Fig 3C. GHR1 was used as a plasma membrane marker. PEPC was used as a cytoplasm marker. The numbers represent the relative ratios of the signal intensity between Myc-SOS2 and GHR1 bands (Myc/GHR1). Statistical analysis of the relative ratios is shown"
NCBI gene IDs:
"Myc: 
pldα1: 820816
pldδ: 829733
SOS2: 833502"
Figure legend: "D SOS2K57G has weak activity and cannot be activated by PA under salt stress. Ten-d-old seedlings expressing 35S:6×Myc-SOS2 or 35S:6×Myc-SOS2K57G in Col-0 or pldα1 pldδ-1 background were treated with or without 100 mM NaCl for 12 h. The brightness and contrast (B&amp;C) of the autoradiograph was adjusted to clearly show the activity of SOS2K57G. The numbers below the blots indicate the relative ratios of the signal intensity between the autoradiograph and Myc-SOS2 bands (Autorad/Myc). Statistical analysis of the relative ratios is shown"
NCBI gene IDs:
"Myc: 
pldα1: 820816
pldδ: 829733
SOS2: 833502"