EMBO/soda-nel huggingface dataset:
Role of Bcl-2 family proteins in a non-apoptotic programmed cell death dependent on autophagy genes (2004)
figf5

(a) Enhancement of etoposide-induced non-apoptotic death by overexpression of human Bcl-2 and Bcl-xL in Bax/Bak−/− MEFs in a Beclin 1-dependent manner. Bax/Bak−/− MEFs with silencing of control Bax or Beclin 1 were transfected with the indicated plasmids (1 μg). After 24 h, the transfected cells were incubated with 20 μM etoposide for 24 h, and cell viability was assessed by the CTB assay. Data are shown as mean ± s.d. (n = 3). (b) Bax/Bak−/− MEFs were transiently transfected with 10 μg of siRNA for Bcl-2, Bcl-x, or Bax (negative control) for 24 h. Expression levels of Bcl-2, Bcl-xL and Lamin B (loading control) were analysed by Western blot analysis. (c–e) Inhibition of etoposide-induced autophagy in Bax/Bak−/− cells treated with siRNA for Bcl-x. (c) Bax/Bak−/− MEFs that were transfected with the indicated siRNAs were treated with 20 μM etoposide for 24 h, then examined by phase-contrast microscopy. (d) Bax/Bak−/− MEFs that were transfected with siRNA for Bcl-x and then treated with etoposide for 18 h were analysed by electron microscopy (×8,500). (e) Reduction of punctate GFP–LC3 fluorescence in etoposide-treated Bax/Bak−/− MEFs by silencing of Bcl-x. Bax/Bak−/− MEFs that were transfected with both GFP–LC3 and the indicated siRNAs were incubated without (NT) or with etoposide for 24 h, and then were examined by confocal fluorescent microscopy. The percentage of cells with punctate GFP–LC3 fluorescence was calculated relative to all GFP–LC3-positive cells. n = 4. (f) Inhibition of etoposide-induced cell death by silencing of Bcl-x. Bax/Bak−/− MEFs were transfected with siRNA (circles, siRNA for Bax; squares, siRNA for Bcl-x) were treated with 20 μM etoposide in the absence (open symbols) or presence (closed symbols) of 10 mM 3-MA, and cell viability was assessed by the CTB assay at the indicated times. n = 4. (g) Clonogenicity assay of Bax/Bak−/− MEFs after exposure to etoposide and staurosporine. Bax/Bak−/− MEFs that were transfected with the indicated siRNAs were treated with 20 μM etoposide or 1 μM staurosporine at 24 h. After collection, 2,000 cells were seeded in the normal medium. After one week, colonies were counted. (h, i) Inhibition of APG5–APG12 induction by silencing of Bcl-x or Beclin 1. Bax/Bak−/− MEFs were treated with the indicated siRNAs (10 μg) for 24 h and then incubated with 20 μM etoposide for the indicated times. Expression of APG5–APG12 complex, Beclin 1 and GAPDH/VDAC (loading control) was analysed by Western blot analysis.
ULK1 translocates to mitochondria and phosphorylates FUNDC1 to regulate mitophagy (2014)
Figure 1

ULK1 is upregulated and translocates to fragmented mitochondria in response to mitophagy induced by hypoxia or FCCP

Levels of ULK1 in MEFs exposed to hypoxia (1% O2) for the indicated times.
Quantification of the ratio of ULK1 to actin in (A).
Levels of ULK1 in MEFs treated with FCCP (20 µM) for the indicated times.
Quantification of the ratio of ULK1 to actin in (C).
MEFs were exposed to hypoxia (1% O2) for 12 h or FCCP (20 µM) for 6 h and then fixed by 4% paraformaldehyde. Cells were stained with anti-TIM23 (mouse) and anti-ULK1 (rabbit) primary antibodies, then Alexa Fluor-488-labeled donkey anti-rabbit IgG and Alexa Fluor-555-labeled donkey anti-mouse IgG secondary antibodies before analysis by immunofluorescence microscopy. Scale bar, 10 µm.
Immunoblots of subcellular fractions from control MEFs or MEFs that were exposed to hypoxia (1% O2) for 12 h. PNS, post-nuclear supernatant; Cyto, cytosol; Mito, mitochondria.
Immunoblots of subcellular fractions from control MEFs or MEFs that were treated with FCCP (20 µM) for 6 h. PNS, post-nuclear supernatant; Cyto, cytosol; Mito, mitochondria.

Data information: All results are from three independent experiments. All quantitative data are presented as mean ± s.e.m. from 3 independent experiments; *** P < 0.001.Source data are available online for this figure.
ULK1 translocates to mitochondria and phosphorylates FUNDC1 to regulate mitophagy (2014)
Figure 2

The mitophagy receptor FUNDC1 is a novel substrate of ULK1 and FUNDC1 (N118A) that prevents its binding to ULK1, impairs ULK1 translocation to mitochondria, and inhibits mitophagy

A HeLa cells were co-transfected with FUNDC1-Myc and FLAG-ULK1. Cells were lysed and immunoprecipitated with anti-Myc antibody 24 h after transfection.
B FLAG-FUNDC1 was co-transfected with GFP-vector or GFP-ULK1 for 24 h in HeLa cells, and then, cell lysates were immunoprecipitated with anti-GFP antibody.
C MEFs were exposed to hypoxia (1% O2) for 12 h or FCCP (20 µM) for 3 h, followed by immunoprecipitation using anti-ULK1 antibody.
D The expression of FUNDC1 is induced by 10 ng/ml tetracycline (Tet) in FUNDC1-inducible HeLa cells. Cells were transfected with HA-ULK1 or HA-ULK1 (K46N) for 24 h. Then, the cell lysates were prepared for immunoblotting using indicated antibodies.
E ULK1+/+ cells were transfected with FUNDC1-Myc, and ULK1 cells were transfected with FUNDC1-Myc alone, FUNDC1-Myc and HA-ULK1, or FUNDC1-Myc and HA-ULK1 (K46N). Cells were harvested and lysed for immunoblots 24 h after transfection.
F Purified GST-FUNDC1 and GST-FUNDC1 (S17A) were subjected to an in vitro kinase assay with Myc-ULK1 immunoprecipitated by anti-Myc antibody from Myc-ULK1-transfected cells about 24 h post-transfection. Phosphorylated FUNDC1 was detected by anti-p-S17-FUNDC1 antibody. The red asterisks mark the target bands.
G Western blot analysis of the kinetics of ULK1, FUNDC1, and the phosphorylated FUNDC1 in MEFs exposed to hypoxia (1% O2) for the indicated times.
H Western blot analysis of the kinetics of ULK1, FUNDC1, and the phosphorylated FUNDC1 in MEFs treated with FCCP (20 µM) for the indicated times.
I, J The single mutant which can abolish the ULK1 and FUNDC1 interaction was identified. HeLa cells were co-transfected with Flag-ULK1 and the indicated constructs. 24 h after transfection, cells were lysed for immunoprecipitation by anti-Flag antibody.
K MEFs were transfected with FUNDC1-Myc and its mutants after the endogenous FUNDC1 was knocked down by siRNA. 36 h after transfection, cells were harvested in the absence or presence of 50 nM bafilomycin A1 (BAF1) and then lysed for immunoblotting with the indicated antibodies.
L MEFs were co-transfected with FUNDC1-Myc or its mutants and MitoDsred after the endogenous FUNDC1 was knocked down by siRNA. 24 h post-transfection, cells were treated with hypoxic (1% O2) conditions for 12 h or FCCP (20 µM) for 6 h before fixed by 4% paraformaldehyde and stained with indicated antibodies. Scale bar, 10 µm.

Data information: All results are from three independent experiments.Source data are available online for this figure.
ULK1 translocates to mitochondria and phosphorylates FUNDC1 to regulate mitophagy (2014)
Figure 3

Mitophagy is restored in ULK1-null MEFs upon reconstitution of the kinase-active form of ULK1 or constitutively active FUNDC1 (S17D)

ULK1+/+ and ULK1-/- cells were cultured under hypoxic (1% O2) condition for 12 h or 24 h in the absence or presence of 50 nM bafilomycin A1 (BAF1). Cell lysates were immunoblotted.
ULK1+/+ and ULK1-/- cells were treated with FCCP (20 µM) for the indicated times in the absence or presence of 50 nM bafilomycin A1 (BAF1). Cell lysates were immunoblotted.
ULK1+/+ and ULK1-/- cells were transfected with FUNDC1-Myc for 24 h and then harvested in the absence or presence of 50 nM bafilomycin A1 (BAF1). Cell lysates were immunoblotted.
ULK1-/- cells were transfected with HA-ULK1 or HA-ULK1 (K46N), then cultured under hypoxic (1% O2) conditions for 24 h in the absence or presence of 50 nM bafilomycin A1 (BAF1). Cells lysates were immunoblotted.
ULK1-/- cells were transfected with HA-ULK1 or HA-ULK1 (K46N) and then treated with FCCP (20 µM) for 24 h. Cells were harvested in the absence or presence of 50 nM bafilomycin A1 (BAF1). Cell lysates were immunoblotted.
FUNDC1-knockdown MEFs were transfected with FUNDC1-Myc or FUNDC1-Myc (S17A) for the indicated times. Cell lysates were used for immunoblotting.
ULK1+/+ cells were transfected with FUNDC1-Myc, and ULK1-/- cells were transfected with FUNDC1-Myc or FUNDC1-Myc (S17D). 24 h after transfection, cells were fixed with 4% paraformaldehyde and stained with anti-Myc (mouse) and anti-LC3 (rabbit) primary antibodies for immunofluorescence microscopy. Scale bar, 10 µm.
ULK1-/- cells were transfected with FUNDC1-Myc or FUNDC1-Myc (S17D) for 24 h in the absence or presence of 50 nm bafilomycin A1 (BAF1) for an additional 6 h before harvesting. Cell lysates were immunoblotted.
ULK1-/- cells were transfected with FUNDC1-Myc or FUNDC1-Myc (S17D) for 24 h in the absence or presence of 50 nm bafilomycin A1 (BAF1) for an additional 6 h before harvesting. Cell lysates were detected with the citrate synthase activity kit.
HeLa cells were transfected with FUNDC1-Myc, FUNDC1-Myc (S17A), or FUNDC1-Myc (S17D). 24 h after transfection, cell lysates were immunoprecipitated with anti-LC3 antibody.

Data information: All results are from three independent experiments. All quantitative data are presented as mean ± s.e.m. from 3 independent experiments; *** P < 0.001.Source data are available online for this figure.
ULK1 translocates to mitochondria and phosphorylates FUNDC1 to regulate mitophagy (2014)
Figure 4

FUNDC1 and ULK1 cooperatively regulate mitophagy

A ULK1+/+ or ULK1-/- cells were cultured under hypoxic (1% O2) or normoxic conditions in the absence or presence of 50 nM bafilomycin A1 (BAF1). ULK1-/- cells were transfected with HA-ULK1, and ULK1-/- FUNDC1-KD cells were transfected with HA-ULK1 and FUNDC1-Myc, then cultured under hypoxic (1% O2) conditions for 24 h in the absence or presence of 50 nM bafilomycin A1 (BAF1). Cells were harvested and lysed for immunoblotting assays.
B ULK1+/+ or ULK1-/- cells were treated with or without FCCP (20 µM) in the absence or presence of 50 nM bafilomycin A1 (BAF1). ULK1-/- cells were transfected with HA-ULK1 and ULK1-/- FUNDC1-KD cells were transfected with HA-ULK1 and FUNDC1-Myc and then treated with FCCP (20 µM) for 24 h in the absence or presence of 50 nM bafilomycin A1 (BAF1). Cells were harvested and lysed for immunoblotting assays.
C ULK1+/+, ULK1-/- FUNDC1-KD, or ULK1-/- FUNDC1-KD cells rescued by introduction of HA-ULK1 and FUNDC1-Myc were cultured under hypoxic (1% O2) conditions for 24 h or with FCCP (20 µM) for 24 h. Samples were analyzed by electron microscopy. Scale bar, 2 µm. The yellow asterisks mark mitochondria. The red arrows indicate double-membraned autophagic structures. The yellow arrow denotes autolysosomes.
D–E The mitochondrial coverage and mitochondrial-containing AVs shown in (C) were quantified. Recon, reconstitution.
F Provisional working model of the interplay between ULK1 and FUNDC1 in mitophagy regulation.

Data information: All results are from three independent experiments. All quantitative data are presented as mean ± s.e.m. from 3 independent experiments; ***P < 0.001.Source data are available online for this figure.
The genomic landscape of nasopharyngeal carcinoma (2014)
figf2

Data matrix showing frequent SCNVs, significantly mutated genes (bold; Benjamini-Hochberg FDR 0.2 calculated with CHASM) and their associated pathway or family mutations discovered in cases with NPC analyzed by WES, targeted deep sequencing (TS) or SNP array (SNP-a). The frequencies of the alterations are plotted on the right. The colors and shapes denoting different types of somatic events are also applied in Figure 5a. Columns, examined cases; rows, genes; hot spot, identical mutations have been registered in COSMIC (see URLs); LOH, loss of heterozygosity. The asterisk indicates that this tumor was also subjected to SNP array profiling. PIK3CA amplification in C666 cells has been previously described.
The genomic landscape of nasopharyngeal carcinoma (2014)
figf4

(a–d) NPC cells stably expressing either negative control shRNA (Scramble) or pooled shRNAs targeting ARID1A (shARID1A) were subjected to anchorage-independent colony formation assay (a), migration assay (b), xenograft growth assay (horizontal lines, mean values) and western blot (WB) assay with the indicated antibodies (d). (e–g) NPC cells either with or without exogenously expressing ARID1A were examined by anchorage-independent colony formation assay (e), migration assay (f) and WB assay with the indicated antibodies (g). (h,i) NPC cells either with or without exogenously expressing BAP1 were examined by anchorage-independent colony formation assay (h) and WB assay with indicated antibodies (i). β-actin was examined as a loading control. The data shown in a, b, e, f and h represent the mean ± s.d.; n = 3 for a, b, e and f; n = 4 for h. *P 0.05, **P 0.01. Student's t-test was used to calculate statistical significance in a–c, e, f and h; the blots shown in d, g and i are representative of three total blots run.
The genomic landscape of nasopharyngeal carcinoma (2014)
figf5

(a) Data matrix showing alterations of genes involved in the ERBB-PI3K signaling pathway and their related targeting molecules. Considering space limitations, the numbers of targeting agents available for each protein are shown in parentheses. Drugs currently in phase 2 clinical trials are marked in blue, those in phase 3 are in purple, and FDA-approved drugs are in brown (see URLs). (b) Kaplan-Meier survival curves of NPC subjects either with or without genetic lesions in the ERBB-PI3K signaling pathway. Log-rank test was used to calculate statistical significance.
LAG3 is not expressed in human and murine neurons and does not modulate α-synucleinopathies (2021)
Figure 1

Fig. 1. Absence of expression of LAG3 in human brain cells.

A. Binding of eight commercial antibodies to recombinant human LAG323-450 and murine LAG324-442 via indirect ELISA. Seven out of eight antibodies bound either human or mouse LAG3, while one antibody (LSB15026) recognized both species.

B. Specific detection of murine but not human LAG3 using 4-10-C9 anti-LAG3 antibody is confirmed with Western Blotting.

C. No detection of human LAG3 in neuronal or glial cell lines of human origin. The band for LAG3 was detected in activated T cells.

D. No band for human LAG3 could be detected with Western Blot in lysates of fully differentiated human NSC-derived neural cultures.

E. Violin plot showing the RNA expression levels of human LAG3 in human NSC-derived neural cultures. Identities annotate different clusters: Neuronal clusters are comprised of the following markers: GAD2, GABRG1, NTRK2, NEFM, SNCG, SLC17A6, SCN2A, DDIT3/HRK. Mixed glial clusters are defind by the following markers: GFAP, S100B, STMN2, NRN1, GPM6B, COL1A1, with astrocyte-specific clusters characterised by GFAP, S100B, GPM6B, COL1A1. LAG3 cannot be evidenced in any of the clusters beyond few random events. Data shown from 5'476 unique analysed cells from one out of two independent biological replicates.

F. Dopaminergic neuronal cultures from control lines and glucocerebrosidase (GBA) N370S PD patients were immunoblotted for the presence of LAG3. No band for LAG3 could be observed in neurons.

G. Using high power, high resolution laser scanning confocal microscopy, no human LAG3 signal could be detected in human neurons (Auto-hLAG3 transduced, DOX OFF) by two different anti-human LAG3 antibodies (17B4 and D2G40; left panel and zoomed-in insets) whereas LAG3 was clearly detected in human neurons induced to express hLAG3 (DOX ON; right panel and zoomed-in insets). Scale bars 25 µm.

H. Human brain homogenates from autopsy material were immunoblotted for the presence of LAG3. No band for LAG3 could be evidenced in any of the brain homogenates. Control samples (tonsils and activated human T cells) show expected bands.

I. Nuclei were isolated from 21 human dorsolateral prefrontal cortices from 16 donors, isolated, and were subjected to snRNAseq. Expression levels were quantified and are shown as violin plots. The LAG3 transcripts are non-detectable in all 34 distinct cell types, including multiple excitatory and inhibitory neurons, oligodendrocytes (ODC), oligodendrocyte precursor cells (OPCs) microglia (MGL), astrocytes (AST), or endothelial cells (EC). Cluster markers as detailed in Saez-Atienzar et al. 2021.
LAG3 is not expressed in human and murine neurons and does not modulate α-synucleinopathies (2021)
Figure 2

Fig. 2. Absence of expression of LAG3 in mouse brain cells, particularly in neurons.

A. Brain homogenates from LAG3 KO and WT mice were immunoblotted for the presence of murine LAG3. Western Blot does not show a specific band other than for activated murine T cells.

B. Mixed neuroglial, neuron-enriched, and glial cortical cultures were assayed for the presence of LAG3 where no band could be found. Activated T cells display band at expected size.

C. Lysates of synaptoneurosomes (SNS) of different brain regions and mouse brain region-specific areas as well as murine activated T cells were assayed for the presence of LAG3 with sandwich ELISA. No evidence for LAG3 could be found in any of the samples other than in activated T cells. Expression is shown in pg LAG3 per µg total protein as mean values of technical quadruplicates with 95% confidence intervals.

D. Using high power, high resolution laser scanning confocal microscopy, no mouse LAG3 signal could be detected in murine primary neuronal cultures (Auto-mLAG3 transduced, DOX OFF and non-transduced neurons) using anti-mouse LAG3 antibody (MABF954) whereas LAG3 was clearly detected in primary neurons induced to express mLAG3 (Auto-mLAG3 transduced, DOX ON). Scale bars 20 µm.

E. RT-qPCR-derived CT values to measure the cDNA copy number. Mixed neuroglial cultures, glial cultures, and neuron-enriched cultures show transcript levels that are close to indistinguishable to those from LAG3‑/‑ BH. Conversely, around 1600 transcripts/ng of RNA were counted in activated T cells.

F. Murine mesencephalon and striatum of 1-year old mice (n = 17 in total) were interrogated with scRNAseq. Violin plot showing expression values of Lag3, in 76'305 assigned cells, between identified cell clusters, mainly expressed in TMEM119-positive microglia cells. Cluster markers are detailed in Russo et al. 2019.
LAG3 is not expressed in human and murine neurons and does not modulate α-synucleinopathies (2021)
Figure 3

Fig. 3. Characterization of binding properties between α-synuclein PFFs and LAG3 reveals promiscuous character of α-synuclein.

A. and B. Binding of ASC filaments (A) or α-synuclein PFFs (B) to multiple proteins. Serial dilutions of ASC filaments and α-synuclein PFFs were performed and bound fractions were detected using anti-ASC or anti-α-synuclein (MJFR1) antibodies. ASC filaments did not interact with any of the proteins (A). Conversely, α-synuclein PFFs strongly bound LAG3, CD4, and MTBD tau and showed moderate binding properties to PrPC, APOE3, and TDP-43. No binding between α-synuclein PFFs and nAra h 2 or BSA could be detected. Values represent means ± SD of technical duplicates.

C. and D. Diffusional sizing of α-synuclein PFFs with fluorescently conjugated LAG3, CD4, BSA, and MJFR1 α-synuclein antibody (C) and of fluorescently labelled LAG3 with monomeric α-synuclein, α-synuclein PFFs, FGL1, and Relatlimab anti-LAG3 antibody (D). An increase in the hydrodynamic radius (Rh) of α-synuclein PFFs is seen with increasing concentrations for MJFR1 but not for LAG3 or any of the control proteins, also not at a lower scale (see insert) (C). Similarly, the hydrodynamic radius of labelled LAG3 increases with higher concentrations of Relatlimab and FGL1 but does not change with α-synuclein PFFs or monomeric α-synuclein (D). Values are given as means ± SD of technical triplicates.

E. and F. Co-immunoprecipitation of LAG3 and subsequent immunoblotting with anti α-synuclein antibody (MJFR1) (E) and co-immunoprecipitation of α-synuclein (monomeric or PFFs) and subsequent immunoblotting with anti-LAG3 antibody (D2G40) (F). Using the anti-α-synuclein antibody for pulldown resulted in LAG3 bands with both monomeric as well as fibrillar α-synuclein. The usage of anti-LAG3 for pulldown led to the specific detection of α-synuclein PFFs, although the PFF fraction predominates also in the input and supernatant. Bands for α-synuclein (E) and LAG3 (F) are indicated by arrows.
LAG3 is not expressed in human and murine neurons and does not modulate α-synucleinopathies (2021)
Figure 4

Fig. 4. Propagation of α-synuclein PFFs in vitro is not dependent on LAG3 in human NSC-derived neural cultures.

Human neural cultures transduced by Auto-hLAG3 were treated by α-synuclein PFFs 4 days post LAG3 expression induction by DOX and kept in culture for 2 or 4 weeks. Both transgenic (hLAG3 D2G40-positive in (A) and 17B4-positive in (B)) and non-transduced, wild-type neurons (selected neurons in zoomed-in insets) propagated α-synuclein and developed characteristic pS129-positive (81A-positive in (A) and EP1536Y-positive in (B)) α-synuclein aggregates. Scale bars 25 µm. Trained ilastik algorithms were used to segment pixels of 81A, EP1536Y and MAP2 stainings imaged by high-content wide-field microscope, which were used to quantify the signal of 81A-positive (C) and EP1536Y-positive (D) α-synuclein aggregates expressed as % of MAP2-positive area. Almost the entire wells (182 fields per well) were imaged for every condition and replicate and each datapoint in the plot represents the entire well. Error bars indicate mean ± SD of biological replicates (duplicates). For few conditions, unicates were used, shown as one dot. One-way ANOVA followed by Tukey's multiple comparison test demonstrated that neurons that did not express LAG3 (DOX OFF) showed no difference in α-synuclein propagation when compared to LAG3-expressing neurons (DOX ON) as demonstrated by two different pS129 α-synuclein antibodies (p=0.9998 for 81A at 2 weeks and p=0.2522 at 4 weeks; p=0.9986 for EP1536Y at 2 weeks and p=0.3042 at 4 weeks).
LAG3 is not expressed in human and murine neurons and does not modulate α-synucleinopathies (2021)
Figure 5

Fig. 5. Survival in A53T α-synuclein mice and aggregation of α-synuclein in vivo and in hippocampal slice cultures is independent from LAG3 expression.

A. Survival curves of A53T α-synuclein TG mice, with LAG3 knockout, heterozygous LAG3 expression, or LAG3 WT. Median survival was shortest for α-synuclein TG LAG3‑/‑ at 239 d, followed by α-synuclein TG LAG3+/- at 260 d and α-synuclein TG LAG3+/+ at 268 d. The survivals of ASYNA53T LAG3‑/‑, LAG3+/- and LAG3+/+ mice were similar (Mantel-Cox log-rank test, p-value = 0.165). Mice were euthanized and dissected in the late-stage symptomatic phase, which would inevitably result in death within a week. None of the control LAG3‑/‑ α-synuclein WT mice died over the course of the experiment.

B. Immunostaining for hyperphosphorylated α-synuclein aggregates (pS129) and aggregated protein (Thioflavin S) in sections of end-stage symptomatic mice; no obvious difference in terms of staining pattern or number of stained cells were apparent in midbrain or brain stem. Scale bars represent 1 mm (sagittal section) and 100 µm (panel).

C. Immunofluorescence staining and quantification of pS129-positive α-synuclein inclusions in hippocampal slice cultures (HSCs) at 5 weeks post-seeding with 350 µM PFF. Scale bars represent 500 µm (overview), and 100 µm (insets). Shown is mean ± SEM and individual values are shown; n = 6 cultures per group; two-way-ANOVA revealed for LAG3 F(1, 20) = 0.0294; p = 0.8656; A53T F(1, 20) = 36.42, p < 0.0001; interaction F(1, 20) = 0.0062, p = 0.9382. Bonferroni's correction for multiple comparisons revealed p = 0.9973 for LAG3 WT vs. KO in A53T TG and p > 0.9808 for LAG3 WT vs. KO in A53T WT.

D. Immunofluorescence staining and quantification of Thioflavin S-positive α-synuclein inclusions in HSCs at 5 weeks post-seeding. Scale bars represent 500 µm (overview), and 100 µm (insets). Shown is mean ± SEM and individual values are shown; n = 5-6 cultures per group; two-way-ANOVA revealed for LAG3 F(1, 19) = 1.972, p = 0.1763; A53T F(1, 19) = 62.49, p < 0.0001; interaction F(1, 19) = 1.586, p = 0.2231. Bonferroni's correction for multiple comparisons revealed p = 0.1373 for LAG3 WT vs. KO in A53T TG and p > 0.9999 for LAG3 WT vs. KO in A53T WT.

EV Figures