EMBO/SourceData huggingface dataset:
Figure legend: "c, d. Incidence of  papilloma  formation (ns: non-significant; Gehan-Breslow-Wilcoxon test) (c) and number of tumours per mouse (d) in wild-type (WT, n=17) and PRSS35-/- (n=22) mice treated with DMBA-TPA (*p=0.0171; Wilcoxon matched-paired rank test). Data represent means ± S.E.M."
NCBI gene IDs:
"PRSS35: 244954"
Figure legend: "b. Relative mRNA expression levels of the indicated  ECM genes in primary  fibroblast  cultures (col1a1: collagen type 1 alpha 1; αSMA, alpha smooth muscle actin;  Cyr61: Cysteine-rich angiogenic inducer 61 (=CCN1)) in primary fibroblast  cultures isolated from WT and PRSS35-/- mice, untreated or after 24 hours stimulation with 10 ng/mL TGFβ1  (n≥7 biological replicates per condition) (** p&lt;0.01, *** p&lt;0.001, **** p&lt;0.0001.; Two-way ANOVA with multiple comparisons). Data represent means ± S.E.M. c Relative  mRNA expression levels of the indicated cytokines/growth factors in primary fibroblast  cultures (n≥8 per condition) (VEGF-α, vascular endothelial growth factor-alpha; FGF2: basic fibroblast growth factor;  FGF7: keratinocyte growth factor) (* p&lt;0.05, ** p&lt;0.01, *** p&lt;0.001, **** p&lt;0.0001.; Two-way ANOVA with multiple comparisons). Data represent means ± S.E.M. "
NCBI gene IDs:
"alpha smooth muscle actin: 11475
αSMA: 11475
Cyr61: 16007
Cysteine-rich angiogenic inducer 61: 16007
col1a1: 12842
collagen type 1 alpha 1: 12842
basic fibroblast growth factor: 14173
FGF2: 14173
FGF7: 14178
keratinocyte growth factor: 14178
PRSS35: 244954
vascular endothelial growth factor-alpha: 22339
VEGF-α: 22339"
Figure legend: "d.  Picrosirius red staining of normal  skin, day 14 post-wounding (d14pw) skin or wound-induced papillomas from WT and PRSS35 KO mice. Red colouring indicates thick  fibres, green colouring indicates thin fibres. Scale bars: 200 µm. e. Quantification of picrosirius red staining as total collagen  density (upper panel) and ratio of red versus green picrosirius staining (lower panel). Using QuPath Bioimage analysis software pixels colouring red or green were determined per area. (n≥4 technical replicates per condition) (* p&lt;0.05, ** p&lt;0.01, **** p&lt;0.0001.; Two-way ANOVA with multiple comparisons). Data represent means ± S.E.M. "
NCBI gene IDs:
"PRSS35: 244954"
Figure legend: "(d) Two distinct siRNAs against NLRP3 and an irrelevant control siRNA were utilized to knock down RNA levels by lipofectamine transfection in macrophages that were later incubated with iRBCL, or RBCL as control, with or without XO, before quantification of secreted IL1β. Results are representative of 2 independent experiments with different donors."
NCBI gene IDs:
"NLRP3: 114548"
Figure legend: "Schematic, western blot, and immunofluorescence  analysis of the 3×FLAG-UBXD8  knockin U2OS  cell line. Scale bar, 5 μm. Data information: In C, mitochondria  were visualized by anti-Tom20 immunostaining; ER  was labeled by anti-PDI immunostaining."
NCBI gene IDs:
"FLAG: 
UBXD8: 23197"
Figure legend: "Blue native gel analysis of UBXD8  in mitochondrial  fractions purified from WT and ∆UBXD8 HeLa  cells."
NCBI gene IDs:
"UBXD8: 23197"
Figure legend: "Immunoprecipitation  analysis of UBXD8 interaction  with mitochondrial  (MARCH5  and MUL1) and ER  (RNF185) ubiquitin E3  ligases. HeLa  cells were infected with lentivirus  to stably express HA-tagged ubiquitin E3  ligases. Whole cell lysates were prepared for anti-HA immunoprecipitation."
NCBI gene IDs:
"HA: "
Figure legend: "Immunoprecipitation  analysis of UBXD8 interaction  with MARCH5  and RNF185. Whole cell lysates from 3×FLAG-UBXD8  knockin HEK293T  cells were prepared for anti-FLAG immunoprecipitation. MARCH5  and RNF185: short exposure for whole cell lysate blots, long exposure for FLAG-IP blots."
NCBI gene IDs:
"FLAG: 
UBXD8: 23197"
Figure legend: "Fractionation analysis of UBXD8's effect on VCP  recruitment to mitochondria  and the ER. Whole cell lysate, mitochondria, and ER  fractions were purified from WT, ∆UBXD8, and ∆UBXD8 HeLa  cells rescued with UBXD8  or the UBX* mutant. UBX*-UBXD8 carries four point mutations (K367A, F407A, P408G, R409A) in the UBX domain to disrupt the UBXD8-VCP interaction. For each sample, 5 μg proteins were loaded for  western blot  analysis."
NCBI gene IDs:
"UBXD8: 23197"
Figure legend: "A, B. Western blot (A) and quantitative (B) analyses of MiD49  and Mcl1  degradation in WT and ∆MARCH5 HeLa  cells. Cycloheximide  (CHX: 200 μg/ml; protein synthesis inhibitor); MG132: 20 μM, proteasome  inhibitor. Data information: Data are shown as mean ± SE from three biological repeats (B, Statistics: two-tailed unpaired Student's t-test (B, ; *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001, n.s., not significant."
NCBI gene IDs:
"MARCH5: 54708"
Figure legend: "Fractionation analysis of the subcellular localization  of mito-UBXD8  and ER-UBXD8. Whole cell lysate, mitochondria, and ER  fractions were purified from the indicated HeLa  cells. Exogenously-expressed mito-UBXD8   has similar expression  level with endogenous UBXD8  in WT cells; exogenously-expressed WT UBXD8  and ER-UBXD8  have similar expression  levels."
NCBI gene IDs:
"UBXD8: 23197"
Figure legend: "D, E. Western blot (D) and quantitative (E) analyses of Mcl1  degradation in the indicated HeLa  cells. ∆UBXD8 HeLa  cells were rescued with vector, WT-, mito-, and ER-UBXD8. Data are shown as mean ± SE from three biological repeats (E). Statistics: two-tailed unpaired Student's t-test (E); *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001."
NCBI gene IDs:
"UBXD8: 23197"
Figure legend: "Western blot analysis of BH3-domain containing proteins in the indicated HeLa  cells treated with Doxo  (10 μM). Three proteins with enhanced level in ∆UBXD8 cells (Noxa, Bik, and Bnip3) are highlighted in red."
NCBI gene IDs:
"UBXD8: 23197"
Figure legend: "C. Localization of GFP fused with wild-type or mutant MLS of GASZ in HeLa cells. Mitochondrial localization was monitored by MitoTracker (Red) staining. Scale bars: 15 m."
NCBI gene IDs:
"GASZ: 74068"
Figure legend: "G. Co-IP assays of GFP-GASZ with various FLAG-tagged wild-type GASZ or its deletion mutants in 293T cells"
NCBI gene IDs:
"GASZ: 74068"
Figure legend: "A. Mitochondria visualized by MitoTracker (Red) when wild-type GASZ or GASZ with MLS mutation was expressed in HeLa cells. Scale bar: 15 m."
NCBI gene IDs:
"GASZ: 74068"
Figure legend: "B. EM on 293T cells with empty vector control (Ctrl), wild-type GASZ, or GASZ with F to P mutation in MLS. Scale bar: 1 m. N: nucleus."
NCBI gene IDs:
"GASZ: 74068"
Figure legend: "C. Averaged percentage of HeLa cells (determined from 100 cells in each experiment) with different types of mitochondria determined by MitoTracker (Red) staining when transfected with GASZ, its SAM/ZIP deletion mutant, or GASZ with MLS mutation."
NCBI gene IDs:
"GASZ: 74068"
Figure legend: "D. HeLa cells transfected with COX4-DsRed, mito-PAGFP, and empty vector control, GASZ or GASZ with MLS mutation, followed by photoactivation of PAGFP. Representative images of PAGFP and DsRed were taken at 0, 10, and 20 min post photoactivation. Scale Bar: 10 m. Insets show images with higher magnification. The averaged exponential decay constant (±s.e.m.) of normalized PAGFP fluorescence within the designated region (white circle) at photoactivation area over a period of 10 min was shown as a bar graph below. n&gt;20 cells for each group."
NCBI gene IDs:
"GASZ: 74068"
Figure legend: "E. Averaged percentage of C18-4 cells (determined from 100 cells in experiment) with different types of mitochondria determined by MitoTracker (Red) staining when transfected with Su9-GFP or GFP-GASZ."
NCBI gene IDs:
"GASZ: 74068
Su9: 66152"
Figure legend: "F. Real-time PCR analyses of mtDNA copy numbers on testes at different time points postnatal, relative to a single copy gene, PECAM, encoded by nuclear DNA. n=5 per genotype."
NCBI gene IDs:
"PECAM: "
Figure legend: "C. Co-IP assays of HA-tagged MFN1 with various FLAG-tagged wild-type GASZ, its domain deletion mutants, or GASZ with MLS mutations in 293T cells."
NCBI gene IDs:
"GASZ: 74068"
Figure legend: "D. Averaged percentage of MEFs with different types of mitochondria were determined according to the mitochondrial morphology by MitoTracker (Red) staining in wild-type MEFs, or MEFs containing MFN null mutations transfected with empty vector control (Ctrl) or a GASZ expressing plasmid."
NCBI gene IDs:
"GASZ: 74068"
Figure legend: "E. PEG fusion assays of Su9-GFP and COX4-DsRed labeled wild-type MEFs, or MEFs containing MFN null mutations transfected with empty vector control (Ctrl) or a GASZ expressing plasmid. Bar graph represents averaged percentage of areas with different levels of mitochondrial fusion."
NCBI gene IDs:
"GASZ: 74068"
Figure legend: "A. Gross morphology of testes from MFN1f/f-Cre compared to their wild-type littermates."
NCBI gene IDs:
"Cre: 
MFN1: 67414"
Figure legend: "B. Histological studies of testis sections from wild-type mice or MFN1f/f-Cre littermates at different time points postnatal. Red stars: tubules with spermatocytes."
NCBI gene IDs:
"Cre: 
MFN1: 67414"
Figure legend: "D. EM of testical sections from wild-type neonatal mice or MFN1f/f-Cre littermates. N: nucleus."
NCBI gene IDs:
"Cre: 
MFN1: 67414"
Figure legend: "B. Whole exome sequencing was carried out and analysis revealed a missense p.L165F variant (c.493 C&gt;T) in the ALX1 homeodomain, heterozygous in the parents (ALX1165L/165F), wildtype in the unaffected sibling (ALX1165L/165L), and homozygous in both affected subjects (ALX1165F/165F)."
NCBI gene IDs:
"ALX1: 8092"
Figure legend: "F. Expression of pluripotent (OCT4, NANOG), endoderm (Endo., AFP, GATA4, FOXA2), ectoderm (Ecto., NESTIN, GFAP, SOX1), and mesoderm (Meso., BRACH. (BRACHYURY), RUNX1, CD34) gene markers for ALX1165L/165L (green), ALX1165L/165F(red), and ALX1165F/165F (blue) iPSC relative to undifferentiated cells (UND). Data are represented as pooled mean ± SEM of three experiments on three clones from each genotype. Significance: p=0.0167 for OCT4, p=0.0005 for NANOG, p= 0.000004 for AFP, p=0.0082 for GATA4, p=0.0137 for FOXA2, p=0.00002 for NESTIN, p=0.0167 for GFAP, p=0.0014 for SOX1, p=0.0117 for BRACHYURY, p=0.0008 for RUNX1 and p=0.0068 for CD34 when comparing undifferentiated and differentiated ALX1165L/165L iPSC. p=0.0013 for OCT4, p=0.0011 for NANOG, p=0.0000003 for AFP, p=0.0003 for GATA4, p=0.0063 for FOXA2, p=0.0001 for NESTIN, p=0.027 for GFAP, p =0.000002 for SOX1, p=0.000009 for BRACHYURY, p=3e-9 for RUNX1 and p=0.000006 for CD34 when comparing undifferentiated and differentiated ALX1165F/165L iPSC. p=0.0201 for OCT4, p=0.006 for NANOG, p=1x10-12 for AFP, p=5x10-13 for GATA4, p=0.0031 for FOXA2, p=0.0292 for NESTIN, p=0.00001 for GFAP, p=6x10-7 for SOX1, p=0.0204 for BRACHYURY, p=0.0009 for RUNX1 and p=0.000003 for CD34 when comparing undifferentiated and differentiated ALX1165F/165F iPSC. Data from each clone were pooled and the mathematical mean was calculated. SEM was used to determine the standard error. To test statistical significance, an ANOVA test was performed. A p-value &lt;0.05 was considered to be statistically significant."
NCBI gene IDs:
"AFP: 174
ALX1: 8092
CD34: 947
FOXA2: 3170
GATA4: 2626
GFAP: 2670
NANOG: 79923
NESTIN: 10763
OCT4: 5460
RUNX1: 861
SOX1: 6656
BRACH: 6862
BRACHYURY: 6862"
Figure legend: "Gene expression analysis across NCC differentiation of unaffected control ALX1165L/165L (green), heterozygous ALX1165L/165F (magenta) and homozygous ALX1165F/165F iPSC: ALX1, neural plate border specifier genes ZIC1, PAX7, PAX3, MSX1, MSX2, DLX5; neural crest specifier genes FOXD3, P75, TFAP2A, SNAI2, TWIST1; and lineage specifier gene HAND2. The RT-qPCR relative expression values were normalized to RPLP0 and GAPDH expression. Data is represented as pooled mean ± SEM of three experiments on three clones from each genotype. Exact p-values are provided in Table EV1. Data from each clone were pooled and the mathematical mean was calculated. SEM was used to determine the standard error. To test statistical significance, an ANOVA test was performed. A p-value &lt;0.05 was considered to be statistically significant."
NCBI gene IDs:
"GAPDH: 
RPLP0: 
ALX1: 8092
DLX5: 1749
FOXD3: 27022
HAND2: 9464
MSX1: 4487
MSX2: 4488
P75: 4804
PAX3: 5077
PAX7: 5081
SNAI2: 6591
TFAP2A: 7020
TWIST1: 7291
ZIC1: 7545"
Figure legend: "A. Homozygous ALX1165F/165F NCC (blue) showed an increase in sensitivity to apoptosis when compared to control ALX1165L/165L NCC (black). The data on the left represents the mean percentage of Annexin V positive cells, indicative of apoptosis, as determined by FACS analysis, with the data on the right being an example of one such experiment. Apoptosis was induced by immersion in a 55°C water bath for ten minutes. Representative experiment for each condition is shown. Data is represented as pooled mean ± SEM of three independent experiments. Data obtained of each clone from three independent experiments were pooled and the mathematical mean was calculated. SEM was used to determine the standard error. To test statistical significance, an ANOVA test was performed. A p-value &lt;0.05 was considered to be statistically significant. . *: Significantly different from the basal apoptosis rate: p=3x10-12 between control NCC basal apoptosis and induced apoptosis, and between ALX1165F/165F NCC basal apoptosis and induced apoptosis. **: Significantly different from control NCC (p=0.0004). "
NCBI gene IDs:
"ALX1: 8092"
Figure legend: "B. Expression levels of cyclins CCNA2 (blue) and CCND1 (orange) in NCC at passages 2 and 3 of ALX1165L/165L and ALX1165F/165F NCC. The RT-qPCR relative expression values were normalized to RPLP0 and GAPDH expression. Data is represented as pooled mean ± SEM of three experiments on three clones from each genotype. *: Significantly different from control NCC at passage 2 (p=0.001 between control and ALX1165F/165F NCC at passage 2 for CCNA2, p=0.0052 for CCND1). **: Significantly different from control NCC at passage 3 (p=0.0494 between control and ALX1165F/165F NCC for CCNA2, p=0.0008 for CCND1)."
NCBI gene IDs:
"GAPDH: 
RPLP0: 
ALX1: 8092
CCNA2: 890
CCND1: 595"
Figure legend: "C. Fluorescence Activated Cell Sorting (FACS) experiments showed that control ALX1165L/165L NCC (green) exhibited increased expression of mesenchymal markers CD90, CD105, and CD73 with culture time (passages 1 through 4), whereas homozygous ALX1165F/165F NCC (blue) showed a consistent expression of the markers expressed at passage 1 throughout. Further, control ALX1165L/165L NCC showed a downregulation of CD57 expression with culture time, while ALX1165F/165F NCC maintained the same level of CD57 across passages. Data are presented as the mean percentage of positive-stained cells across passage numbers. Data obtained of each clone from three independent experiments were pooled and the mathematical mean was calculated. SEM was used to determine the standard error. To test statistical significance, an ANOVA test was performed. A p-value &lt;0.05 was considered to be statistically significant. *: Significantly different from control NCC. For CD90, p=0.0013 at passage 3 and p=0.0207 at passage 4. For CD105, p=0.0016 at passage 2, p=0.00004 at passage 3 and p=0.0021 at passage 4. For CD73, p=0.0060 at passage 2, p=0.00004 at passage 3 and p=0.0114 at passage 4. For CD57, p=0.0026 at passage 2, p=0.000003 at passage 3 and p=0.000007 at passage 4."
NCBI gene IDs:
"ALX1: 8092"
Figure legend: "A. Mutant ALX1165F/165F NCC (blue) exhibited a migration defect in timed coverage of the central clearing of the wound assay when compared with control ALX1165L/165L NCC (black). Data is presented as percent area recovery of the central circular clear area of the wound assay by migrating NCC at the end of a 24-hour period. For fluorescent pictures, cells were stained in serum free media containing 3.6µM CellTracker Green CMFDA (Life Technologies) for 30 min at 37°C and allowed to recover for 30 min before starting the experiment. Images were acquired every 6 hours using a Keyence BZ-X800 microscope. Surface area analyses and percentages of coverage were measured using ImageJ software (NIH). The NCC were monitored over 24 hours. The data are represented as the average of the percentage of closure ± SEM. Scale bar = 200µm. Data obtained of each clone from three independent experiments were pooled and the mathematical mean was calculated. SEM was used to determine the standard error. To test statistical significance, an ANOVA test was performed. A p-value &lt;0.05 was considered to be statistically significant.*: Significantly different from control NCC (p&lt;0.0001)."
NCBI gene IDs:
"ALX1: 8092"
Figure legend: "B. Multiplex analysis of BMP2 and BMP9 in the supernatant of cultured NCC showed that ALX1165F/165F NCC (blue) secrete less BMP2 and more BMP9 compared to control ALX1165L/165L NCC (green). Data are represented as pooled mean ± SEM of three clones from each genotype. Statistical significance was determined with an ANOVA test. A p-value &lt;0.05 was considered significant. *: Significantly different from control NCC (p =0.0424 for BMP2 and p=0.0192 for BMP9)."
NCBI gene IDs:
"ALX1: 8092"
Figure legend: "C. Addition of soluble BMP2 or CV2, a BMP9 antagonist, to the culture medium could partially rescue the migration defect of ALX1165F/165F NCC. At the beginning of the assay, 10, 50, or 100 ng/ml of soluble BMP2, CV2, or a combination of the two at 100 ng/ml each were added to the culture medium, and the cells were monitored over the next 24 hours. The data are represented as the average of the percentage of closure ± SEM. Scale bar: 400µm."
NCBI gene IDs:
"ALX1: 8092"
Figure legend: "A. Dissected flatmount wild type and alx1-/- zebrafish larvae craniofacial cartilages after Alcian blue staining, the anterior points to the left of the page in all images. The ventral cartilages appear normal, but the alx1-/- anterior neurocranium (ANC) appears narrow, with the midline element that is derived from the frontonasal NCC being absent. The Meckel's cartilage (arrow, MC) is also diminutive. Scale bar: 200 µm."
NCBI gene IDs:
"alx1: 565176"
Figure legend: "B. Zebrafish alx1 mutants (blue) show reduced detectable expression of alx1 but increased expression of alx3, alx4a compared to wild type controls (green). alx4b expression levels are similar between wild type and alx1-/- lines. Data is represented as the mean of all pooled embryos from three different clutches. The RT-qPCR relative expression values were normalized to elfa and 18S expression using the ΔΔCT method. Data from each clutch were pooled and the mathematical mean was calculated. SEM was used to determine the standard error. To test statistical significance, an ANOVA test was performed. A p-value &lt;0.05 was considered to be statistically significant. Statistical significance denoted by *; p&lt;0.0001 between WT zebrafish and alx1-/- at all measured time points; at 10 ss, 24 hpf and 36 hpf for alx3; and at 24 hpf and 36 hpf for alx4a. Refer to Table EV2 for p-values."
NCBI gene IDs:
"18S: 
elfa: 
alx1: 565176
alx3: 566955
alx4a: 100006399
alx4b: 497424"
Figure legend: "C. Dissected flatmount of zebrafish embryos injected with Alx1DN, after Alcian blue staining. The embryos developed an absence of the frontonasal derived median portion of the anterior neurocranium (ANC) and a profound hypoplasia of the Meckel's and ventral cartilages. In the most severely affected zebrafish, a nearly abrogated ANC was observed. Scale bar: 200 µm."
NCBI gene IDs:
"Alx1: 565176"
Figure legend: "D. Lineage tracing experiments in control and Alx1DN mutant embryos revealed aberrant migration of anterior cranial NCC when alx1 is disrupted. In the control animal, the anterior cranial NCC always migrate to contribute to the median portion of the ANC. In contrast, the anterior cranial NCC labeled in the Alx1DN animals fail to migrate to the median ANC, where the ANC structure is narrower and the labeled cranial NCC are found in an anterior and lateral ectopic location (white asterisks). Scale bar: 250µm."
NCBI gene IDs:
"alx1: 565176
Alx1: 565176"
Figure legend: " (B) Bar diagram representation of the relative SUMO conjugation activity of Nse2, full-length Smc5-Nse2, ΔHinge/Smc5-Nse2, ΔHead/Smc5-Nse2 and Arm/Smc5-Nse2 truncation constructs (schematic representation above). Orange bars indicate the presence of ssDNA (virion ϕx174) and red bars absence of ssDNA. Reaction rates were performed at least in three different independent experiments (see Fig EV2). Data values are mean ± s.e.m.; and n=3 technical replicates. Significance was measured by a two-tailed unpaired t-test relative to wild-type. **P&lt;0.01 "
NCBI gene IDs:
"Nse2: 856695
Smc5: 854123"
Figure legend: " (C) SYPRO-stained (left) and Western blot (right) time course SUMO conjugation reaction using and ΔHead/Smc5-Nse2 truncation construct in the presence of ssDNA. T7-tagged Smc5 and E1 were immunodetected by an anti-T7 antibody. Reactions were run at 30 °C and stopped at indicated times by adding SDS-loading buffer "
NCBI gene IDs:
"Nse2: 856695
Smc5: 854123"
Figure legend: " (D) SYPRO-stained (left) and Western blot (right) time course SUMO conjugation reaction using and ΔHinge/Smc5-Nse2 truncation construct in the presence of ssDNA. T7-tagged Smc5 and E1 were immunodetected by an anti-T7 antibody. Reactions were run at 30 °C and stopped at indicated times by adding SDS-loading buffer "
NCBI gene IDs:
"Nse2: 856695
Smc5: 854123"
Figure legend: " (E) Western blot of the time-course SUMO conjugation reaction in the presence of ssDNA (virion ϕx174) using either Nse2 (left) or Arm/Smc5-Nse2 complex (right). The reactions were run in the presence or absence of cNse4 external substrate. Reaction was run at 30°C and stopped at indicated minutes by adding SDS-loading buffer (N-S2, cNse4-SUMO2; N-2S2, cNse4-2SUMO2; N-3S2, cNse4-3SUMO2 and pS2, poly-SUMO2) "
NCBI gene IDs:
"Nse2: 856695
Smc5: 854123"
Figure legend: " (D) Bar diagram representation of the SUMO conjugation rates of activity assays of Arm/Smc5-Nse2 KE mutants in the presence (orange bars) or absence (red bars) of ssDNA (virion ϕx174), relative to wild-type (set to 1). Reactions rates were performed at least in three different independent experiments. Data values are mean ± s.e.m. and n=3 technical replicates. Significance was measured by a two-tailed unpaired t-test relative to wild-type. *P&lt;0.05, **P&lt;0.01, ***P&lt;0.001 "
NCBI gene IDs:
"Nse2: 856695
Smc5: 854123"
Figure legend: " (C) Degree of conformational changes in CM induced by 60b linear ssDNA. Blue bar, Arm/Smc5-Nse2 (wild-type); grey bar, Arm/Smc5-Nse2 (K333E/K344E); green bar, Arm/Smc5-Nse2 (K764E); pink bar, Arm/Smc5-Nse2 (K333E/K344E/K764E); red bar, Arm/Smc5-Nse2 (K743E/K745E). Reactions were performed at least in three different independent experiments. Data values are mean ± s.d. and n=3 technical replicates. Significance was measured by a two-tailed unpaired t-test relative to wild-type. ****P &lt; 0.0001 "
NCBI gene IDs:
"Nse2: 856695
Smc5: 854123"
Figure legend: " (B) Growth test analysis of wild-type, nse2-CH and the indicated smc5-KE mutants; 10-fold serial dilutions of the liquid cultures were spotted in YPD and pictures taken after 48 hours "
NCBI gene IDs:
"nse2: 856695
smc5: 854123"
Figure legend: " (C) Same as in (B) but using the nse2-CH, smc5-3KE, smc5-K743,745E, smc5-K743,745R, smc5-7KE and smc5-7KR mutants "
NCBI gene IDs:
"nse2: 856695
smc5: 854123"
Figure legend: " (A) Protein extracts from exponentially growing wild-type, smc5-3KE, smc5-K743,745E, smc5-4KE and smc5-K743,745R cells were prepared under denaturing conditions; 6xHis-Flag tagged SUMO (HF-SUMO) was pulled down from protein extracts to purify sumoylated species and analyzed by western blot. A strain with no HF-tag was used as control; arrow points to unmodified form of the protein, vertical bar to sumoylated forms "
NCBI gene IDs:
"smc5: 854123"
Figure legend: " (B) Left, same as in (A), but using wild-type and two independent clones of smc5-7KE; right, same as in (A), but using wild-type, smc5-7KE and smc5-7KR mutant cells "
NCBI gene IDs:
"smc5: 854123"