EMBO/soda-nel huggingface dataset:
PAQR3 controls autophagy by integrating AMPK signaling to enhance ATG14L-associated PI3K activity (2016)
Figure 6

Figure 6. Phosphorylation of PAQR3 T32 is required for autophagy initiation upon glucose starvation.

(A) Bacterial purified His-tagged WT or T32A NH2-terminal 71aa of PAQR3 was incubated with or without Flag-tagged ATG14L purified from HEK293T cells. Then the complexes were incubated with AMPK for the indicated time and subjected to Phos-tag gel or regular SDS-PAGE.

(B) PAQR3-deficient HeLa cells were transfected with WT or T32A mutants of PAQR3 plasmids. 24 h later, the cells were incubated with normal medium (NM) or glucose starvation (GS) for 4 h, followed by immunoblotting (IB) in Phos-tag gel or regular SDS-PAGE.

(C) HeLa Cells were incubated with NM or GS for 1 h or treated with compound C (C.C, 20 mM, 30 min) prior to GS for 1 h. In parallel, 25 mM 2-DG or 1 mM AICAR was added in NM for 1 h. Cell lysates were analyzed by IB.

(D) HEK293T cells were transfected with Myc-tagged PAQR3. 24 h after transfection, the cells were treated with GS for 4 h. Then the cell lysates were used in IB and immunoprecipitation (IP) with the antibodies as indicated. The immunoprecipitates were treated with CIP to remove phosphate groups and analyzed by IB in Phos-tag gel or regular SDS-PAGE.

(E) WT, phosphorylation defective (T32A), or phospho-mimetic (T32E) PAQR3 were transfected in the presence or absence of constitutively active AMPK (CA-AMPK) into PAQR3-deficient HeLa cells as indicated. 24 h later, the cells without AMPK transfection were treated with GS for 4 h. 10% of the cell lysates were subjected to IB to detect PAQR3 expression level (shown in Appendix Fig S4A), the remaining cell lysates were used to immunoprecipitate VPS34 complexes by ATG14L antibody, followed by VPS34 activity measurement by a quantitative PI(3)P ELISA assay. The PI(3)P level was normalized to the amount of ATG14L (n = 5; *p < 0.05; **p < 0.01 as compared to the first group).

(F) PAQR3-deficient HeLa cells were infected with lentivirus expressing WT or T32A PAQR3. Then PAQR3 expression levels were examined by IB.

(G) Both WT HeLa cells and PAQR3 knockout HeLa cells infected with lentivirus expressing WT or T32A PAQR3 were treated with GS for 2 h or 4 h respectively. Cell lysates were analyzed by IB.
PAQR3 controls autophagy by integrating AMPK signaling to enhance ATG14L-associated PI3K activity (2016)
Figure 7

Figure 7. PAQR3-deleted mice have deficiency in exercise-induced autophagy.

(A, B) Representative immunohistochemical images of LC3 staining in skeletal muscle (A) and liver (B) after 80 min exercise in WT and PAQR3-deleted mice. Scale bar in (A): 20 μm. Scale bar in (B): 50 μm.

(C, D) Immunoblotting analysis of skeletal muscle (C) and liver (D) from WT or PAQR3 knockout mice at rest or after 80 min exercise.

(E) Abnormal limb-clasping of PAQR3 knockout mice when suspended by its tail. Quantification scoring of the limb clasping phenotype is shown on the right (n = 7 for each group; **p < 0.01).

(F) Motor performance was measured using an accelerating rotarod apparatus, and the time of each mouse spent on the rotating rod until it fell off was recorded. Average time on the rod for each group is shown (n = 7 for each group; **p < 0.01).

(G) Co-immunoprecipitation of Beclin1 or ATG14L with their binding partners in liver tissues from mice of indicated genotype. Immunoprecipitation (IP) and immunoblotting (IB) were performed using the antibodies as indicated.

(H) The supernatants of liver lysates form WT or PAQR3 knockout mice were subjected to gel filtration analysis, followed by IB.

(I) VPS34 complexes immunopurified with Beclin1 or ATG14L antibody in mouse liver were subjected to a quantitative PI(3)P ELISA assay (n = 5; **p < 0.01).

(J) A schematic model of PAQR3 regulation on autophagy in response to glucose starvation.
Estrogen receptor alpha signaling in dendritic cells modulates autoimmune disease phenotype in mice (2022)
Figure 1

Figure 1. Endogenous ERα signal is crucial for determining EAE disease phenotype in drug sensitivity.

(A) EAE severity in 6-to-8-week-old female (left panel) and male (right panel) wild-type (WT) and ERα-/- mice (n=7-9 mice per group).

(B) EAE sensitivity to IFNβ in 6-to-8-week-old female (top panel) and male (bottom panel) WT and ERα-/- mice. Vehicle (Veh) or IFNβ (47 μg/kg) were i.p. injected. every other day from 0 to 10 dpi (n= 8-12 mice per group).

Data information: Data are presented as mean ± SEM. Statistical significance was assessed by an unpaired Student's t-test. *P<0.05
Estrogen receptor alpha signaling in dendritic cells modulates autoimmune disease phenotype in mice (2022)
Figure 2

Figure 2. Interferon-β is ineffective against EAE in mice that lack extra-gonadal estrogen.

(A) Sensitivity of EAE to IFNβ in female wild-type (WT) and Cyp19-/- mice. Vehicle (Veh) or IFNβ (47 μg/kg) were i.p. injected every other day from 0 to 10 dpi (n=5-7 mice per group).

(B) Sensitivity of EAE to IFNβ in ovariectomized (OVX) female mice (n=5-6 mice per group).

(C) Schematic for ovary transplantation experiment.

(D) Sensitivity of EAE to IFNβ in Cyp19-/- ovary-transferred female WT mice and WT ovary-transferred Cyp19-/- mice (n=5-6 mice).

(E) Estradiol levels in LNs (A: axillary, B: branchial, C: cervical, and I: inguinal) in sham-operated and OVX mice (n=5 mice per group).

Data information: Data are presented as mean ± SEM. Statistical significance was assessed by an unpaired Student's t-test. *P<0.05.
Estrogen receptor alpha signaling in dendritic cells modulates autoimmune disease phenotype in mice (2022)
Figure 3

Figure 3. Degree of neuronal damage in the spinal cord is dependent on ERα expression.

(A) Brightfield micrograph of ventral horns of Golgi-Cox stained lumber (L)5 spinal cord section of male WT and ERα-/- EAE mice at 33 dpi with quantitation of area stained in gray matter. The white dotted line indicates the division between white and gray matter, V.H. represents the ventral horn (n=5 mice per group). Scale bar= 50 μm.

(B) Neurite outgrowth of the neurons in ventral horns of Golgi-Cox stained L5 spinal cord section of male WT and ERα-/- mice at 33 dpi with quantitation using Sholl analysis method. Sholl rings are separated by 10 μm (n=4 mice per group, 20 neurons analyzed per animal) with an end radius of 100 μm.

(C) Representative confocal reflection maximum intensity projection micrograph of Golgi-Cox stained dendrites in the lumbar ventral horn of male WT and ERα-/- mice at 33 dpi with quantitative analysis of dendrite spine density (n=4-5 mice per group, 20 dendrites analyzed per animal). Scale bar= 5 μm.

Data information: Data are presented as mean ± SEM. Statistical significance was assessed by an unpaired Student's t-test. *P<0.05.
Estrogen receptor alpha signaling in dendritic cells modulates autoimmune disease phenotype in mice (2022)
Figure 4

Figure 4. T cells derived from EAE-ERα-/- mice are neurotoxic and induce severe and prolonged EAE disease.

(A) Upregulated and downregulated genes in RNAseq analysis were evaluated by qPCR. Splenic CD4+ T cells from male WT EAE and ERα-/- EAE mice at 9 dpi were used (n=5-8 mice).

(B) Schematic for intrathecal passive EAE model. CD4+ T cells were isolated from the spleen and LNs of male WT or ERα-/- EAE mice at 9 dpi. Isolated CD4+T cells were adoptively transferred by intrathecal injection to Tcra-/- recipient mice that were induced with EAE (14 dpi). EAE scores were measured for 20 days. In another group, Tcra-/- recipient mice were sacrificed 14 days after i.th. injection.

(C) EAE scores after CD4+T cell transfer. Tcra-/- mice treated with vehicle (10 µl PBS with 2% FBS) (□), and transferred WT EAE mice- (○) or ERα-/- EAE mice (●)-derived CD4+T cells (3x106 cells in 10 µl). n=5.

(D) Neurite outgrowth of the neurons in Golgi-stained spinal sections of Tcra-/- recipient mice that received passive intrathecal injections of CD4+ splenic cells from male WT EAE and ERα-/- EAE mice at 14 dpi quantified using the Sholl technique. Sholl rings are separated by 10 μm (n=4 mice per group, 20 neurons analyzed per animal) with an end radius of 100 μm.

Data information: Data are presented as mean ± SEM. Statistical significance was assessed by an unpaired Student's t-test. **P<0.01, *P<0.05.
Estrogen receptor alpha signaling in dendritic cells modulates autoimmune disease phenotype in mice (2022)
Figure 5

Figure 5. Dendritic cell ERα is key in mediating T cell neurotoxicity and disease drug sensitivity.

(A) Sensitivity of EAE to IFNβ in 6-to-8-week-old female (top panel) and male (bottom panel) ERαfl/fl (WT) and CD11c cre-ERα-/- (cERα-/-) mice. Vehicle or IFNβ (47 μg/kg) were i.p. injected every other day from 0 to 10 dpi (n= 8 mice per group).

(B) EAE scores after CD4+T cell transfer isolated from cERα-/- EAE at 9 dpi. The detail method is described in Fig. 4B.

(C) Neurite outgrowth of the neurons in Golgi-stained spinal sections of Tcra-/- recipient mice that received passive intrathecal injections of CD4+ splenic cells from male WT EAE and cERα-/- EAE mice at 14 dpi quantified using the Sholl analysis method. Sholl rings are separated by 10 μm (n=4 mice per group, 20 neurons analyzed per animal) with an end radius of 100 μm.

Data information: Data are presented as mean ± SEM. Statistical significance was assessed by an unpaired Student's t-test. **P<0.01, *P<0.05.
Estrogen receptor alpha signaling in dendritic cells modulates autoimmune disease phenotype in mice (2022)
Figure 6

Figure 6. ERα negatively regulates lymphotoxin production in dendritic cell.

(A) Number of membrane-bound lymphotoxin (mLT)+ DCs (derived from spleen and LNs) in male cERα-/- EAE mice at 9 dpi (n=5 mice per group). DC population were gated as Ly6G- and CD11c+ (EV Fig.1 A).

(B) Tuj1-stained differentiated N2a cells co-cultured with nothing, non-treated CD4+ T cell, and rLT-treated CD4+ T cell (10 ng/ml) for 24 h. Panels depict representative fields for each condition. Quantification of percentage differentiated neuron and average neurite length. The results are derived from at least six biological samples. Scale bar= 10 μm.

(C) Vehicle or LTβR-blocking Fc (7.5 μg/kg) were i.p. administered every other day from 0 to 10 dpi (n=8 mice per group).

Data information: Data are presented as mean ± SEM. Statistical significance was assessed by an unpaired Student's t-test (A, C) and Tukey's multiple comparison test following one-way ANOVA (B) . **P<0.01, *P<0.05.
Estrogen receptor alpha signaling in dendritic cells modulates autoimmune disease phenotype in mice (2022)
Figure 7

Figure 7. ERα activation suppresses lymphotoxin expression in dendritic cells.

(A) Lta mRNA expression in BMDCs developed from bone marrows of WT and ERα-/- mice 3 h after treatment with LPS (100 ng/ml) with vehicle (Veh) or PPT (1 µM) pre-treatment for 18 h (n=4 samples per group).

(B) Il1b and Ifnb mRNA expression 3 h after treatment with LPS (100 ng/ml) with vehicle (Veh) or PPT (1 µM) pre-treatment for 18 h (n=4-5 samples per group).

(C) Representative Western blot of TRAF3 and HSC70 in WT BMDCs and ERα-/- BMDCs. TRAF3 expression in BMDCs following vehicle (Veh) or PPT (1 µM) pre-treatment for 18 h was evaluated by normalization of HSC70 (n=5-7 per group).

(D) Representative Western blot of co-immunoprecipitation of ERα and TRAF3 in BMDCs 0, 3, or 6 h after PPT (1 µM) treatment. Ubiquitination of TRAF3 in BMDCs following PPT (1 µM) treatment for 0, 3, or 6 h. Western blot of HSP70 in pre-IP sample as a control.

(E) Representative Western blotting images and quantification assay of phospho-IRF3, IRF3, and GAPDH in BMDCs treated with vehicle or with LPS (100 ng/ml) for 2 h prior upon a vehicle or PPT (1 µM) pre-treatment for 18 h. The results are presented as a ratio of the phospho-IRF3 signal to the IRF3 signal (n=5 samples).

(F) Nuclear localization of IRF3 in BMDCs treated with vehicle or with LPS for 2 h prior upon a vehicle or PPT (1 µM) pre-treatment for 18 h. The results are presented as a ratio of the nuclear IRF3 signal to the cytosol IRF3 signal (n=5-9 biological samples) Scale bar= 20 μm.

(G) ChIP assay for IRF3 bindings to Lta and Ifnb promoter using BMDCs treated with vehicle or with LPS (100 ng/ml) for 2 h prior to a vehicle or PPT (1 µM) pre-treatment for 18 h (n=5 samples).

Data information: Data are presented as mean ± SEM. Statistical significance was assessed by Tukey's multiple comparison test following one-way ANOVA (A, B, E, F, G) and an unpaired Student's t-test (C). **P<0.01, *P<0.05.

Expanded View Figure Legends
Rapid movement and transcriptional re-localization of human cohesin on DNA (2016)
Figure 1

Figure 1. Recombinant human tetrameric cohesin complexes bind to DNA, translocate rapidly in high salt buffer and are released following DNA or cohesin cleavage.

(a) Instant Blue stained SDS-polyacrylamide gel of Scc1wt cohesin tetramers used in (b). (b) Cohesin loading assay. Scc1wt cohesin tetramer was incubated with nicked circular (CN) or 1x, 2x or 4x concentration of linearised (L) plasmid DNA, immunoprecipitated with anti-Scc1 antibodies, washed with high salt buffer and then eluted with Scc1 peptide. Recovered DNA was separated by agarose gel electrophoresis and stained with GelRed DNA stain. Input DNA = 7 %. (c) Silver stained SDS-polyacrylamide gel of non-cleaved and cleaved Scc1Halo-TEV cohesin tetramers used in (d). (d) Non-cleaved and cleaved Scc1Halo-TEV cohesin tetramer was incubated with nicked circular plasmid DNA and processed as in (b). Input DNA = 4 %. (e) Kymograph of Scc1GFP-TEV cohesin bound to doubly tethered λ-DNA in cohesin binding buffer and washed with 750 mM NaCl buffer + Sytox Orange. XhoI flow in induced DNA cleavage and cohesin release. Flow in from top and scale bar = 5 µm in this and all subsequent kymographs. (f) Kymographs of Scc1GFP-TEV cohesin and Scc1GFP cohesin bound to doubly tethered λ-DNA and washed with 750 mM NaCl buffer. TEV protease flow in released Scc1GFP-TEV but not Scc1GFP from DNA.
Rapid movement and transcriptional re-localization of human cohesin on DNA (2016)
Figure 2

Figure 2. Single cohesin complexes bind to DNA and translocate rapidly in high salt buffer.

(a) Kymograph of a single DNA-bound Scc1Halo-TMR-TEV cohesin complex after 750 mM NaCl buffer wash. (b) Distribution of background subtracted EMCCD counts of Scc1Halo-TMR-TEV cohesin complexes immobilized on the coverslip. Peaks at ~ 4 x 103 and ~ 8 x 103 EMCCD counts correspond to single fluorophores and a small fraction of double fluorophores respectively. n = 290 regions with fluorescent molecules, 228 regions with background. (c) Photobleaching kinetics of a single DNA-bound Scc1Halo-TMR-TEV cohesin complex. (d) Diffusion coefficients of Smc1/3 wild type or K38A ATP binding deficient ‘KA’ forms of Scc1Halo-TMR-TEV-cohesin in the presence or absence of ATP. Diffusion coefficients were calculated based on the linear fit of the average mean square displacement of ≥ 13 freely diffusing molecules per condition over a 1 second time period.
Rapid movement and transcriptional re-localization of human cohesin on DNA (2016)
Figure 3

Figure 3. Cohesin bypasses DNA-bound TetRHalo-TMR, Halo-TMREcoRIE111Q and TMR-labelled nucleosomes but not QDotEcoRIE111Q.

(a – c) Kymograph of salt-resistant Scc1GFP-TEV-cohesin diffusing past (a) TetO-DNA-bound TetRHalo-TMR, (b) DNA-bound Halo-TMREcoRIE111Q and (c) a DNA-bound TMR-labelled nucleosome. (d) Kymograph of salt-resistant Scc1GFP-TEV-cohesin failing to bypass DNA-bound QDotEcoRIE111Q. DNA was stained with Sytox Orange.
Rapid movement and transcriptional re-localization of human cohesin on DNA (2016)
Figure 4

Figure 4. Transcription constrains cohesin translocation.

(a) Kymograph of two Halo-TMRT7 RNAP transcription elongation events. (b, c) Kymographs showing (b) Halo-TMRT7 RNAP transcription stalling following removal of NTPs and (c) subsequent resumption after NTP flow in. (d) Kymograph of Halo-TMRT7 RNAP (diameter ~ 8 nm excluding HaloTag) displacing Scc1GFP-TEV-cohesin following resumption of transcription in T7 reaction buffer + NTPs. (e) Kymograph of Halo-TMRT7 RNAP constraining translocation of salt-resistant Scc1GFP-TEV-cohesin.
Rapid movement and transcriptional re-localization of human cohesin on DNA (2016)
Figure 5

Figure 5. CTCF constrains cohesin translocation.

(a) Representative field of view showing Halo-TMRCTCF bound to pPlat-4xCTCF following 75 mM NaCl, 75 mM KCl buffer wash. Arrows denote CTCF bound at predicted site of 4xCTCF array. (b) Kymograph of salt-resistant Scc1GFP-TEV-cohesin failing to bypass DNA-bound Halo-TMRCTCF. DNA was post-stained with Sytox Green.
Allosteric inhibition of SHP2 uncovers aberrant TLR7 trafficking in aggravating psoriasis (2021)
Figure 1

Figure 1. SHP2 expression increased in both human psoriatic patients and IMQ-induced psoriasis-like mice.

A Expression of PTPN11 (gene encoding SHP2) in skin lesions in psoriatic patients compared with skin from healthy donors based on microarray data (No. GSE14905).

B Expression of ptpn11 in human PBMCs of psoriatic patients (n=14) and normal controls (n=16).

C Representative SHP2 staining in skin sections of psoriatic patients (n=13) and normal controls (n=5). Scale bars: 200 μm.

D Western blot analysis of PBMCs lysates derived from psoriatic patients and normal controls.

E The catalytic activity of SHP2 was measured in human PBMCs lysates derived from psoriatic patients (n=25) and normal controls (n=25).

F Representative p-ERK staining of skin sections of psoriatic patients and normal controls. Scale bars: 200 μm.

G Quantitative PCR analysis of Ptpn11 mRNA levels in the IMQ-treated or non-treated dorsal back of C57BL/6 mice at day 5 (n=6/group). Data were normalized to Gapdh expression.

H Representative histological sections of IMQ-treated or non-treated dorsal back of C57BL/6 mice at day 5. Scale bar: 100 μm.

I Quantitative PCR analysis of mRNA levels in human PBMCs derived from psoriatic patients (n=20) and normal healthy controls (n=20).

Data information: Data are represented mean ± SEM. P values are determined by two-tailed Mann-Whitney U test (A and B) or two-tailed unpaired Student's t-test (E, G, and I). *P<0.05, **P<0.01.
Allosteric inhibition of SHP2 uncovers aberrant TLR7 trafficking in aggravating psoriasis (2021)
Figure 2

Figure 2. SHP2-inhibitor SHP099 attenuated the psoriasis-like phenotype in the IMQ-induced murine model.

C57BL/6 mice (n=6/group) were treated with indicated dose of SHP099 or vehicle for 4 days.

A Phenotypic presentation (top), H&E staining (middle) and statistical data (bottom) (mean ± SEM) of dorsal skin. Scale bar: 100 μm.

B Clinical scores plotted with mean ± SEM. *denotes statistical significance when compared with the IMQ group.

C ELISA quantification of protein levels of cytokines in mouse serum.

D, E Quantitative PCR analysis of mRNA encoding IL-23/IL-17A axis cytokines (D) and other psoriasis-related cytokines (E) in the dorsal skin of C57BL/6 mice treated with indicated dose of SHP099 or vehicle for 4 days. Results were normalized to Gapdh expression.

Data information: Data are represented mean ± SEM. P values are determined by two-tailed unpaired Student's t-test (A, C-E) or Tukey multiple comparison test (B). *P<0.05, **P<0.01.
Allosteric inhibition of SHP2 uncovers aberrant TLR7 trafficking in aggravating psoriasis (2021)
Figure 5

Figure 5. SHP2 deficiency in myeloid cells alleviated the psoriasis-like phenotype in the IMQ-induced murine model.

Representative images (A and E), histological sections of dorsal back (B and F) and clinical scores (C and G) and quantitative PCR analysis of mRNA levels in dorsal skin (D and H) from wild-type (n=6) and M-Shp2-/- mice or DC-Shp2-/- mice (n=6) treated with IMQ for 4 days. Scale bar: 100 μm. Left: H&E (hematoxylin and eosin) staining data; right: statistical data (mean ± SEM). Results were normalized to Gapdh expression.

Data information: Data are represented mean ± SEM. P values were calculated by Tukey multiple comparison test (B, D, F, H) or Bonferroni multiple comparison test (C, G). *P<0.05, **P<0.01, ns, not significant.
Allosteric inhibition of SHP2 uncovers aberrant TLR7 trafficking in aggravating psoriasis (2021)
Figure 6

Figure 6. SHP2 deficiency in macrophages mitigated psoriasis by suppressing NF-κB activation.

A Heat map showing mRNA expression in peritoneal macrophages derived from wild-type and M-Shp2-/- mice after IMQ treatment based on RNA sequencing (n=3/group). Colors represent high (red) and low (blue) intensity. Genes labeled in green indicate they are in the NF-κB signaling.

B Volcano plot image of upregulated (in red) and down regulated (in green) genes (M-Shp2-/- mice compared to wild-type mice) in peritoneal macrophages from wild-type and M-Shp2-/- mice after IMQ treatment (n=3/group).

C Expression levels of representative psoriasis-related genes decreased in the IMQ model of M-Shp2-/- mice compared to wild-type mice (n=3/group).

D BMDMs and PMs derived from wild-type and M-Shp2-/- mice were stimulated by IMQ (10 µg/ml) for indicated times. Whole cell lysates were subjected to Western blotting.

E Representative p-p65 staining of skin sections from mouse. Scale bar: 100 μm.

Data information: Data are represented mean ± SEM. P values are determined by Tukey multiple comparison test (C). **P<0.01, ns, not significant.
Allosteric inhibition of SHP2 uncovers aberrant TLR7 trafficking in aggravating psoriasis (2021)
Figure 7

Figure 7. SHP2 interacted with TLR7 and promoted TLR7 trafficking to endosomes.

A HEK293T cells were transfected with plasmids expressing HA-tagged SHP2 and GFP-tagged TLR7. After 48 h, cells were left unstimulated or stimulated by IMQ (10 µg/ml) for 30 min. Cell lysates were immunoprecipitated by anti-HA or anti-GFP and probed by indicated antibodies.

B PMA-differentiated THP-1 cells were unstimulated or stimulated by IMQ (10 µg/ml) for 30 min. Cell lysates were immunoprecipitated by anti-TLR7 or anti-SHP2 and probed by indicated antibodies.

C BMDMs derived from wild-type mice were stimulated by IMQ (10 µg/ml) for 30 min. Cell lysates were immunoprecipitated by anti-SHP2 and probed by indicated antibodies.

D Representative images of confocal microscopy performed with PMA-differentiated THP-1 cells treated with IMQ (10 µg/ml) for indicated durations. Scale bar: 20 µm.

E Immunoblotting of HEK293T cells co-transfected for 48 h with GFP-TLR7, plus HA-SHP2, HA-SHP2 mutant vectors, followed by immunoprecipitation with anti-GFP beads.

F PMA-differentiated THP-1 cells were infected with vector or SHP2 lentivirus and then treated with or without R848 (10 µg/ml). Subcellular fractionation was performed, and cytoplasmic and cell membrane proteins were probed by respective antibodies.

G, H Immunoblot analysis of TLR7 and SHP2 in the isolated ER, Golgi and endosome from PMA-differentiated THP-1 cells with either SHP2 overexpressed (G) and SHP2 deficiency(H).

I Skin sections from psoriatic patients and healthy controls were immune-stained for TLR7 together with a marker of the early endosome (EEA1) or late endosome/lysosome (LAMP1) prior to analysis by confocal microscopy, showing TLR7 localization in different organelles (arrows). Scale bar: 50 μm.