EMBO/SourceData huggingface dataset:
Figure legend: "Quantification by  real-time qPCR analysis of Il6, Il17a, II22, Ccl3, Ccr6, and Ccl20  mRNAs in whole skin  of naïve and irradiated  (15Gy) control (PBS) and CSA  treated WT mice  at 14 days post-IR, (n = 5). Data information: Data represent mean ± SD. *P &lt; 0.05, **P &lt; 0.01 by two-tailed Mann-Whitney test; two independent experiments."
NCBI gene IDs:
"Ccl20: 20297
Ccl3: 20302
Ccr6: 12458
Il17a: 16171
II22: 50929
Il6: 16193"
Figure legend: "The shRNA targeting TAZ, but not YAP, knocked down the expression of the smaller protein (asterisk)."
NCBI gene IDs:
"TAZ: 6901"
Figure legend: "Exogenous TAZ/YAP was not processed proteolytically to cTAZ. RKO cells were transfected with the indicated plasmids expressing C-terminal HA-tagged TAZ or YAP."
NCBI gene IDs:
"HA: 
TAZ: 
YAP: "
Figure legend: "RKO, but not HEK293A cells, expressed both full-length TAZ and cTAZ transcripts. RT PCR primers targeting different regions"
NCBI gene IDs:
"cTAZ: 
TAZ: 6901"
Figure legend: "Knock out of cTAZ in RKO cells using CRISPR/Cas9 technology. Monoclones (#3 and #14) were selected."
NCBI gene IDs:
"CRISPR: 
cTAZ: 
Cas9: 901176"
Figure legend: "Cluster analysis of RNA expression of WT, cTAZ-/-, and cTAZ-/- /GFP-cTAZ put-back RKO cells. Genes down- and up-regulated more than 2 folds (P &lt; 0.05) in cTAZ KO cells were included in cluster analysis. Green and red indicate down- and up-regulated genes respectively."
NCBI gene IDs:
"cTAZ: 
GFP: "
Figure legend: "cTAZ suppressed expression of ISGs. RNA levels of selected ISGs in WT, cTAZ-/-, cTAZ-/-/GFP-cTAZ RKO cells were quantified by qPCR. Error bars indicate SD, n =3. **P&lt;0.01; ***P&lt;0.001; Student's t-test."
NCBI gene IDs:
"cTAZ: 
GFP: "
Figure legend: "cTAZ repressed 5×ISRE-luciferase activity induced by IFN-α. HEK293A cells were transfected with 5×ISRE-luciferase reporter along with the indicated plasmids, treated with or without IFN-α (50 ng/ml) for 12 hours. The expression of ectopic genes was determined by IB. Error bars indicate SD, n =3. ***P&lt;0.001; one-way ANOVA test was used for statistical analysis."
NCBI gene IDs:
"cTAZ: 
luciferase: "
Figure legend: "Knockout of cTAZ promoted ISGs expression in the presence of IFN-α. WT and cTAZ-/- RKO cells were treated with or without of IFN-α (50 ng/ml) for indicated time, the expression of selected genes was quantified by qPCR. WT or cTAZ-/- group was normalized by basal level, respectively. Error bars indicate SD, n =3. *P&lt;0.05; **P&lt;0.01; Student's t-test."
NCBI gene IDs:
"cTAZ: "
Figure legend: "cTAZ attenuated heterodimerization between STAT1 and STAT2 induced by IFN-α. Following transfection, cells were treated with or without IFN-α (50 ng/ml) for 1 hour and subjected to IP assays using Flag antibody."
NCBI gene IDs:
"cTAZ: "
Figure legend: "Depletion of cTAZ enhanced the interaction between STAT1 and STAT2. RKO cell lines (WT and #14) were treated with IFN-α as in (B). IP assays were performed using STAT1 antibody."
NCBI gene IDs:
"cTAZ: "
Figure legend: "cTAZ attenuated interaction between STAT1 and importin α5 (Im-α5). Following transfection, HEK293A cells were stimulated using IFN-α IP assays were performed using Flag antibody."
NCBI gene IDs:
"cTAZ: "
Figure legend: "Deletion of cTAZ promoted nuclear accumulation of STAT1 upon IFN-α stimulation. RKO (WT and cTAZ-/-) cells treated with or without IFN-α (50 ng/ml) for 1 hour, fixed, and subjected to IF using STAT1 antibody (green). DNA was labeled by DAPI (blue). Scale bar, 50 μm. Quantification was performed using ImageJ software (right). N&gt;C: nuclear; N=C: nuclear plus cytoplasmic; N&lt;C: cytoplasmic localization."
NCBI gene IDs:
"cTAZ: "
Figure legend: "Deletion of cTAZ promoted nuclear accumulation of STAT1 upon IFN-α stimulation. Cells were treated and subjected to subcellular fractionation."
NCBI gene IDs:
"cTAZ: "
Figure legend: "cTAZ defective in STAT1-binding failed to block nuclear translocation of STAT1. HEK293A stably overexpressing STAT1 were transfected with the indicated plasmids, treated with or without IFN-α (50 ng/ml) for 1 hour, fixed, and subjected to IF Scale bar, 25 μm."
NCBI gene IDs:
"cTAZ: 
STAT1: "
Figure legend: "cTAZ repressed the antiviral activity of ectopic STAT1/2. HEK293A were transfected with indicated plasmids, infected with gVSV (MOI = 0.01) for 12 hours, fixed, and subjected to microscopy (Left). Viral infection and replication were marked by green fluorescence. Scale bar, 100 μm. (Right) Quantification of GFP positive cells. About 300 cells (from 3 different fields) were analyzed. Error bars indicate SD. **P&lt;0.01; ***P&lt;0.001; Student's t-test."
NCBI gene IDs:
"cTAZ: 
GFP: 
STAT1: "
Figure legend: "cTAZ effectively repressed antiviral activity of IFN-α. Transfection and infection were performed Top: GFP intensity of cell lysates was measured by a fluorometer. Bottom: viral GFP and ectopic cTAZ protein levels were detected by IB."
NCBI gene IDs:
"cTAZ: "
Figure legend: "Depletion of cTAZ repressed viral replication. RKO cell lines (WT and two independent cTAZ -/- clones) were infected with gVSV (MOI = 0.1) for 12 hours. Scale bar, 100 μm. The ratio of GFP positive cells were calculated About 300 cells (from 3 different fields) were analyzed. Error bars indicate SD. ***P&lt;0.001; Student's t-test."
NCBI gene IDs:
"cTAZ: 
GFP: "
Figure legend: "Depletion of cTAZ repressed viral replication. Cells were treated GFP intensity of cell lysates was measured. Error bars indicate SD, n = 3. **P&lt;0.01; ***P&lt;0.001; Student's t-test."
NCBI gene IDs:
"cTAZ: 
GFP: "
Figure legend: "Depletion of cTAZ repressed viral replication. Cells were treated the viral genomic RNA was quantified by qPCR (normalized to human GAPDH). Error bars indicate SD, n = 3. ***P&lt;0.001; Student's t-test."
NCBI gene IDs:
"cTAZ: 
GAPDH: 2597"
Figure legend: "cTAZ deficiency enhanced ISGs expression. Cells were treated the expression of IFIH1 and DDX58 were assessed by qPCR. Error bars indicate SD, n = 3. *P&lt;0.05; **P&lt;0.01; ***P&lt;0.001; Student's t-test."
NCBI gene IDs:
"cTAZ: 
DDX58: 23586
IFIH1: 64135"
Figure legend: "Overexpression of cTAZ promotes virus infection. DOX-inducible cTAZ-overexpressing RKO cell lines were pre-treated with DOX for 48 hours (1 μg/ml). gVSV infection and GFP Fluorescence intensity quantification were performed Error bars indicate SD, n = 3. **P&lt;0.01; ***P&lt;0.001; Student's t-test."
NCBI gene IDs:
"cTAZ: "
Figure legend: "Overexpression of cTAZ repressed ISGs expression. Cells were treated the expression of IFIH1 and DDX58 were assessed by qPCR. Error bars indicate SD, n = 3. *P&lt;0.05; **P&lt;0.01; Student's t-test."
NCBI gene IDs:
"cTAZ: 
DDX58: 23586
IFIH1: 64135"
Figure legend: "IFN-α induced mRNA level of cTAZ. RKO cells were treated with different doses of IFN-α for 8 hours. RNA levels were measured by qPCR. Error bars indicate SD, n = 3. ***P&lt;0.001; two-way ANOVA test was used for statistical analysis."
NCBI gene IDs:
"cTAZ: "
Figure legend: "STAT1/2 synergistically activated a cTAZ promoter. HEK293A cells were transfected with cTAZ reporter with or without STAT proteins. Promoter activity was determined using luciferase assay, and protein expression was determined by IB. Error bars indicate SD, n = 3. ***P&lt;0.001; Student's t-test."
NCBI gene IDs:
"cTAZ: 
luciferase: 
STAT1: "
Figure legend: "STAT1/2 binding sites on cTAZ promoter. HEK293A were transfected with cTAZ promoters (wild type or mutants). Luciferase activity and responses to STAT1/2 expression were measured. Error bars indicate SD, n = 3. **P&lt;0.01; Student's t-test."
NCBI gene IDs:
"cTAZ: "
Figure legend: "STAT1 occupied cTAZ promoter in an IFN-α-sensitive manner. RKO cells were treated with or without IFN-α (50 ng/ml) for 1 hour and subjected to ChIP assays. cTAZ PCR primers were designed to target STAT1/2 binding site. ISG15 promoter was included as a positive control."
NCBI gene IDs:
"cTAZ: 
ISG15: 9636"
Figure legend: "(B) DraI digestion of PCR products from the multiple organs of the four fetuses (Upper). NC, negative control with no genomic DNA loaded. Mut, MITFL247S/L247S genomic DNA loaded. WT, LW genomic DNA loaded. Multiple organs of the four fetuses were examined for KIT expression by agarose gel electrophoresis (Bottom). Mut, MITFL247S/L247S genomic DNA loaded. WT, LW genomic DNA loaded"
NCBI gene IDs:
"KIT: 396810
MITF: 414902"
Figure legend: "(A) Representative microscopic appearances of the retina and RPE cells (arrowhead) of a chimeric, a WT and a Mut fetus at the same gestational age by H&amp;E staining. WT, Bama WT fetus. Mut, Bama MITFL247S/L247S fetus. NW-2, the chimeric fetus. Scale bars, 100 μm"
NCBI gene IDs:
"MITF: 414902"
Figure legend: "(B) Representative immunofluorescence images showed the expression of multiple RPE markers Pax6, MITF and Bestrophin in the chimeric fetuses. Blue, Hochst3342. WT, Bama WT fetus as positive control. NW-2, the chimeric fetus. MITFL247S/L247S fetus as negative control. Scale bars, 50 μm"
NCBI gene IDs:
"MITF: 414902"
Figure legend: "(B) Digestion of the MITF PCR products in multiple organs of the full-term chimeric pig. NC, negative control with no genomic DNA loaded. Mut, MITFL247S/L247S genomic DNA loaded. WT, Bama GFP-labeled genomic DNA loaded. In the chimera piglet, GFP-specific primers were used to further confirm the chimeric contribution to multiple organs in the piglet. GAPDH was used to confirm the DNA quality of all the samples. NC, negative control with no genomic DNA loaded. Mut, Bama MITFL247S/L247S genomic DNA loaded. WT, Bama GFP-labeled genomic DNA loaded"
NCBI gene IDs:
"MITF: 414902"
Figure legend: "(D) Representative immunofluorescence images showed the expression of a corneal epithelial cell marker (K12) in GFP-labeled cells. WT, Bama WT piglet. Mut, Bama MITFL247S/L247S piglet. CH, chimeric piglet. Scale bars, 50 μm"
NCBI gene IDs:
"MITF: 414902"
Figure legend: "(B) Detection of WASH gene targeting. The floxed WASH gene by gene targeting was analysed by Southern blotting (left panel) and PCR (middle panel). The genotypes of the offspring were analysed by PCR (right panel)."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(C) WASH deficiency is embryonic lethal. E11.5 of WASH deficiency showed a shrinked decidua."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(D) Histology analysis of E7.5 WASH+/+ and WASH−/−embryos. Scale bar, 100 μm."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(E) WASH‐deficient embryos do not undergo apoptosis. TUNEL assay was performed for apoptosis detection. PI, propidium iodide. FITC, fluoresceinisothiocyanate. Scale bar, 100 μm."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(F) E7.5embryos of WASH−/−mice present extensive autophagy. Ultrastructure of isolated E7.5embryos was stained with antibody against LC3 and visualized by immuno‐electron microscopy. Black arrowhead indicates autophagosome. *, Cellular reference. Scale bar, 500 nm (upper panel). WASH+/+ or WASH−/−embryos were isolated, and LC3 conversion and p62 level were analysed by immunoblotting (lower panel). Bands were quantified and analysed by Image J and shown as means±s.d. ***P0.001. Above experiments were repeated for three independent times with similar results.Source data for this figure is available on the online supplementary information page."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(A) WASH deficiency enhances autophagy induction. WASH+/+ or WASH−/− MEFs were treated with EBSS for the indicated times in the presence or absence of 20 nM bafilomycin A1 (BafA1), and harvested for immunoblotting. Ratios of LC3‐II/β‐actin were calculated and shown at the right panel."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(B) WASH knockout accelerates LC3 conversion by confocal microscopy. Endogenous LC3 puncta were visualized by staining with antibody against LC3 (left panel), and calculated as shown in the right panel. Nuclei were stained with DAPI. Scale bar, 10 μm."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(C) Poly‐ubiquitinated proteins are reduced in WASH−/− MEFs. WASH+/+ and WASH−/− MEFs were starved in EBSS for 2 h treated with or without 20 nM BafA1 or 10 μM MG132. Cells were harvested for immunoblotting with anti‐poly‐ubiquitin antibody (Enzo, clone FK1) that only recognizes poly‐ubiquitinated ubiquitin chains."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(D) Autophagosome‐like structures in WASH+/+ and WASH−/− MEFs were visualized after starvation for 1 h by immuno‐electron microscopy with antibody against LC3 (left panel). Black arrowhead indicates autophagosome. Scale bar, 500 nm. *, Cellular reference. Fifty cells were quantified from three independent experiments (right panel). Data were shown as means±s.d. *P0.05 and **P0.01. Experiments were repeated for three independent times with similar results.Source data for this figure is available on the online supplementary information page."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(A) WASH colocalizes with GFP‐LC3 upon starvation. HeLa cells stably expressing GFP‐LC3 were stained with anti‐WASH antibody after treatment with EBSS or culture medium (CM) for 1 h."
NCBI gene IDs:
"LC3: 440738///81631///84557"
Figure legend: "(A) WASH deficiency enhances the interaction of Vps34 with Beclin 1. WASH+/+ and WASH−/− MEFs were treated with EBSS at 37°C for 1 h and immunoprecipitated for Beclin 1. IP, immunoprecipitation. Ratio of Vps34/Beclin 1 was calculated by the Image J software and shown in the right panel."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(B) WASH KO MEFs were treated with or without EBSS at 37°C for EBSS followed by kinase assays. Cells were lysed and incubated with anti‐Beclin 1 antibody to precipitate the autophagy‐related Vps34. Immunoprecipitates were separated into two equal parts, one for kinase assays and the other for input detection. RLU, relative light unit."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(C) WASH deficiency enhances Vps34 activity. WASH+/+ and WASH−/− MEFs stably expressing GFP‐WIPI1 were stimulated with or without EBSS for 1 h followed by confocal microscopy (left panel). GFP‐WIPI1 dots were calculated and shown in the right panel. Scale bar, 10 μm."
NCBI gene IDs:
"WASH: 68767
WIPI1: 52639"
Figure legend: "(D) WASH‐silenced HeLa cells were treated with or without EBSS at 37°C for 1 h, and detected as above."
NCBI gene IDs:
"WASH: 100287171"
Figure legend: "(E) WASH attenuates the interaction of Beclin 1 with Vps34. MEFs stably expressing vector or Flag-WASH were starved with EBSS for 1 h, followed by immunoprecipitation with anti‐Beclin 1 antibody. Ratio of Vps34/Beclin 1 was shown in the right panel."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(F) WASH overexpression reduces Vps34 activity. MEFs stably expressing GFP‐WIPI1 were transfected with vector or Flag-WASH and then starved with EBSS for 1 h, followed by visualization with confocal microscopy (upper panel). GFP‐WIPI1 dots were calculated and shown in the lower panel. Scale bar, 10 μm. Data are shown as means±s.d. **P0.01 and ***P0.001. Experiments were repeated for three independent times with similar results.Source data for this figure is available on the online supplementary information page."
NCBI gene IDs:
"WASH: 68767
WIPI1: 52639"
Figure legend: "(A) The interaction of WASH with Beclin 1 was validated by yeast two‐hybrid assays. Yeast strain AH109 was co‐transfected with Gal4 DNA‐binding domain (BD) fused WASH and Gal4 activating domain (AD) fused Beclin 1. p53 and large T antigen or a known WASH interactor Bloc1s2 was introduced as positive controls."
NCBI gene IDs:
"Gal4: 855828
WASH: 100287171"
Figure legend: "(B, C) WASH co‐immunoprecipitates Beclin 1 in mammalian cells. Flag‐tagged Beclin 1 and Myc‐tagged WASH were co‐transfected into HEK293T cells and immunoprecipitations were performed at 24 h post transfection. IP, immunoprecipitation."
NCBI gene IDs:
"Beclin 1: 8678"
Figure legend: "(E) The indicated Flag‐tagged Beclin 1 constructs encoding different regions of Beclin 1 were co‐transfected with full‐length HA‐tagged WASH (HA-WASH) into HEK293T cells followed by immunoprecipitation."
NCBI gene IDs:
"Beclin 1: 56208"
Figure legend: "(C) Lysine 437 mutation to arginine (K437R) abrogates ubiquitination of Beclin 1. Different constructs of Flag‐tagged Beclin 1 mutants were co‐transfected with HA‐tagged ubiquitin (HA-Ub) into HEK293T cells."
NCBI gene IDs:
"Beclin 1: 8678"
Figure legend: "(D) Beclin 1 (WT) or K437R‐Beclin 1 (K437R) mutant was stably expressed in Beclin 1‐silenced HeLa cells. Beclin 1‐knockdown HeLa cells were infected with lentivirus encoding 3XFlag‐tagged WT Beclin 1 (WT) or K437R‐Beclin 1 (K437R)."
NCBI gene IDs:
"Beclin 1: 8678"
Figure legend: "(E) K437R‐Beclin 1 mutant abrogates the enhanced interaction of Beclin 1 with Vps34 during autophagy. HeLa cells expressing 3XFlag-Beclin 1 or 3XFlag-K437R‐Beclin 1 were treated with EBSS for 1 h, and harvested for immunoprecipitation."
NCBI gene IDs:
"Beclin 1: 8678"
Figure legend: "(F) Vps34 kinase activity is impaired by K437R‐Beclin 1 mutant. HeLa cells were transfected with the indicated vectors for 24 h and immunoprecipitated with anti‐HA antibody followed by a Vps34 kinase assay. Immunoprecipitates were separated into two equal parts, one for kinase assays and the other for input detection."
NCBI gene IDs:
"Beclin 1: 8678"
Figure legend: "(G) K437R‐Beclin 1 reduces the activation of Vps34. Beclin 1‐silenced HeLa cells stably expressing GFP‐HeLa were rescued with WT‐ or K437R‐Beclin 1, followed by stimulation with 1 h for 1 h. GFP‐WIPI1 dots were visualized by confocal microscopy."
NCBI gene IDs:
"Beclin 1: 8678"
Figure legend: "(H) K437R‐Beclin 1 mutant fails to induce autophagy after starvation. Beclin 1‐knockdown HeLa cells expressing WT or K437R mutant of Beclin 1 (K437R) were starved with EBSS for the indicated times."
NCBI gene IDs:
"Beclin 1: 8678"
Figure legend: "(I) K437R‐Beclin 1 mutant suppresses autophagosome formation. Beclin 1‐knockdown HeLa cells expressing WT‐Beclin 1 or K437R‐Beclin 1 were infected with lentivirus encoding GFP‐LC3, and stimulated with EBSS for 1 h in the presence or absence of 20 nM BafA1. GFP‐LC3 dots were visualized by confocal microscopy. Scale bar, 10 μm. Data are shown as means±s.d. **P0.01 and ***P0.001. All the above experiments were repeated for at least three times with similar results.Source data for this figure is available on the online supplementary information page."
NCBI gene IDs:
"Beclin 1: 8678"
Figure legend: "(A) WASH overexpression suppresses the ubiquitination of Beclin 1. HEK293T cells were transfected with the indicated vectors for 24 h followed by an Ni‐NTA‐based pull‐down assay and immunoblotted with anti‐poly‐ubiquitin (anti‐poly‐Ub) (left panel), and anti‐K63‐specific ubiquitin (anti‐K63‐Ub) (middle panel) antibodies. The same blot was stripped and probed with anti‐K48‐specific ubiquitin (anti‐K48‐Ub) antibody (right panel)."
NCBI gene IDs:
"Beclin 1: 8678
WASH: 100287171"
Figure legend: "(B) WASH overexpression hinders K63‐linked ubiquitination of Beclin 1. HeLa cells stably expressing vector or Flag-WASH were treated with CM or EBSS with or without BafA1 for 1 h and then stained with antibodies against endogenous Beclin 1 and K63‐linked polyubiquitin (K63‐Ub). The colocalization rate (Pearson's correlation coefficient) between Beclin 1 and K63‐Ub was calculated and shown in the right panel."
NCBI gene IDs:
"WASH: 100287171"
Figure legend: "(C) WASH overexpression inhibits LC3 lipidation. HeLa cells stably expressing vector or Flag-WASH were treated with CM or EBSS with or without BafA1 for 1 h and then stained with antibody against endogenous LC3. LC3 dots were calculated and shown in the right panel."
NCBI gene IDs:
"WASH: 100287171"
Figure legend: "(D) WASH deficiency increases K63‐linked ubiquitination of Beclin 1. WASH+/+ and WASH−/− MEFs were treated with EBSS at 37°C for 1 h and lysed for immunoprecipitation (IP) with anti‐Beclin 1 antibody and probed with antibody against K63‐linked poly‐ubiquitin."
NCBI gene IDs:
"WASH: 68767"
Figure legend: "(E) WASH overexpression inhibits Vps34 activity. HeLa cells were transfected with the indicated vectors and subjected to immunoprecipitation with anti‐Beclin 1 antibody for Vps34 kinase activity assay. Immunoprecipitates were separated into two equal parts, one for kinase assay and the other for input detection. RLU, relative light unit."
NCBI gene IDs:
"WASH: 100287171"
Figure legend: "(G) Ambra1 knockdown declines Beclin 1 ubiquitination. HeLa cells with Ambra1 knockdown were treated with EBSS for 1 h, followed by immunoprecipitated with anti‐Beclin 1 antibody. Immunoprecipitates were dissociated with 1% SDS and re‐immunoprecipitated with anti‐Beclin 1 antibody, followed by immunoblotting with the indicated antibodies."
NCBI gene IDs:
"Ambra1: 55626"
Figure legend: "(H) Ambra1 overexpression increases Beclin 1 ubiquitination. HeLa cells with Ambra1 overexpression were treated as above."
NCBI gene IDs:
"Ambra1: 55626"
Figure legend: "(J, K) WASH and Ambra1 competitively bind to Beclin 1. HeLa cells with vector or WASH overexpression were starved for 1 h and lysed for immunoprecipitation with anti‐Beclin 1 antibody (J). Recombinant Ambra1 and WASH proteins were incubated with GST-Beclin 1, followed by GST pull‐down assays (K). Scale bar, 10 μm. Data are shown as means±s.d. **P0.01 and ***P0.001. Data were repeated for at least three times with similar results.Source data for this figure is available on the online supplementary information page."
NCBI gene IDs:
"WASH: 100287171"
Figure legend: "D. Mdm12 binds NBD-PE. Wild-type and monomeric (I5P mutant) Mdm12 were incubated with NBD-PE and separated from free NBD-PE in native PAGE. Coomassie staining (left) and fluorescent (right) detection indicates that Mdm12 directly interacts with NBD-PEin vitro.E. Quantitative data showing binding affinities for NBD-PE by Mdm12. The binding affinities of Mdm12 (monomer/dimer shown in Native PAGE and I5P mutant) for NBD-PE was measured with a NBD-PE concentration-dependent manner. All experiments were carried out three times and the means ± SD are given."
NCBI gene IDs:
"Mdm12: 854153"
Figure legend: "F. Mdm12 mutants (L256W, I262W, and L256W/I262W double mutant) were incubated with NBD-PE, and subjected to native PAGE. Because wild-type Mdm12 separates as both monomer and dimer on native PAGE, the purely monomeric form (I5P) of Mdm12 was used as the wild type for clarity. The graph in the right indicates the quantities measured in the experiments. The bar shows the relative amounts of the band ratio (fluorescence/Coomassie). Values represent the means and SD from three independent experiments."
NCBI gene IDs:
"Mdm12: 854153"
Figure legend: "A. Overall structure (left) and electron density (right) of Mdm12 in the asymmetric unit. Four molecules (two Mdm12 dimers) are organized with 2-fold rotation symmetry. The 2-fold axes are indicated with a black dotted line.B. Overall structure (left) and electron density (right) of ΔMdm12 in the asymmetric unit. Six ΔMdm12 molecules (three Mdm12 dimers) are arranged with 2-fold rotation symmetry as shown above."
NCBI gene IDs:
"Mdm12: 854153"
Figure legend: "(C) 293T cells were transfected with empty vector (EV), a vector encoding FLAG-tagged full length CYLD (wt), or constructs encoding FLAG-tagged CYLD fragments 1-581 (F1) lacking the C-terminus, or 581-956 (F2) lacking the N-terminus, as indicated, followed by FLAG-IP. The blot was probed with antibodies recognizing SPATA2, FLAG and tubulin. Endogenous SPATA2 was co-immunoprecipitated to comparable levels by CYLD and the C-terminal CYLD protein fragment, containing the USP domain."
NCBI gene IDs:
"CYLD: 1540"
Figure legend: "(D) CYLD-/- MEF were infected with retrovirus encoding FLAG-CYLD as indicated and CYLD was purified by FLAG-IP. The blot was probed with antibodies recognizing CYLD, SPATA2 and actin."
NCBI gene IDs:
"CYLD: 1540"
Figure legend: "(E) 293T cells were transfected with empty vector (EV), a vector encoding FLAG-tagged full length SPATA2 or a construct encoding the FLAG-tagged N-terminal part of SPATA2 (NT), as indicated, followed by FLAG-IP. The blot was probed with antibodies recognizing CYLD, SPATA2 and tubulin. M, marker lane."
NCBI gene IDs:
"SPATA2: 9825"