EMBO/SourceData huggingface dataset:
Figure legend: " D. PBL34 phosphorylation is induced by 3-OH-C10:0. The protoplast of Col-0 was transformed with 35S:PBL34-FLAG and 35S:PBL34-2A-FLAG, respectively. Sixteen hours later, the protoplasts were treated with1 μM 3-OH-C10:0. Phosphorylated proteins were immunoprecipitated and detected by anti-pS/T antibody. The experiment was repeated three times with similar results. "
NCBI gene IDs:
"35S: 
FLAG: 
PBL34: 831360"
Figure legend: " E. 3-OH-C10:0-induced PBL34 phosphorylation depends on LORE. The 35S:PBL34-FLAG was introduced to Col-0 and lore, respectively. Four-week-old homozygous transgenic plants were treated with 1 μM 3-OH-C10:0. Phosphorylated proteins were immunoprecipitated and detected by anti-pS/T antibody. The experiment was repeated three times with similar results. "
NCBI gene IDs:
"35S: 
FLAG: 
PBL34: 831360
lore: 842432"
Figure legend: " F. PBL34 dissociated from LORE upon 3-OH-C10:0 activation. The 35S:LORE-FLAG plants were transformed with native promoter driven PBL34 (pPBL34:PBL34-HA). 100 nM flg22 or 1 μM 3-OH-C10:0 was sprayed onto plant leaves, and the leaves were sampled for assays15 min later. The experiment was repeated three times with similar results. "
NCBI gene IDs:
"35S: 
FLAG: 
PBL34: 831360
LORE: 842432"
Figure legend: " G. Phosphorylation of PBL34K180E did not disassociate from LORE. The 35S:LORE-FLAG protoplasts were transfected with 35S:PBL34K180E-HA. 100nM flg22 or 1 μM 3-OH-C10:0 was used to treat the protoplasts. The protoplasts were sampled 15 min later. The experiment was repeated three times with similar results. "
NCBI gene IDs:
"35S: 
FLAG: 
PBL34: 831360
LORE: 842432"
Figure legend: " H. PBL34 rescues pbl34/35/36 triple mutant ROS burst in response to 3-OH-C10:0. Treatment was same as in (E). pPBL34:PBL34-HA was introduced to pbl34/35/36 triple mutant, and the homozygous transgenic line was used for ROS burst assays. Values are means ± SD (n=6 biological replicates). (ANOVA, P &lt;0.01). "
NCBI gene IDs:
"pbl34: 831360
PBL34: 831360"
Figure legend: " I. PTI marker gene expression in PBL34 complementation lines. The PTI marker genes FRK1 and NHL10 expression was examined by RT-qPCR in Col-0, pbl34/35/36, and CM-PBL34-2A lines after 3-OH-C10:0 treatments for 4 hrs. Values are means ± SD (n=6; ANOVA; p &lt; 0.01). "
NCBI gene IDs:
"PBL34: 831360
pbl34: 831360
FRK1: 816436
NHL10: 818171"
Figure legend: " J. PBL34 phosphorylation at T306/T310 is essential for disease resistance. pbl34/35/36 triple mutants were complemented with PBL34 or PBL34-2A driven by its native promoter. The homozygous plants were pretreated with 3-OH-C10:0(C10) 1 d before pathogen inoculation. The plants were inoculated with 2×105 cfu/ml Pst DC3000, and the bacterial titer was analyzed at 3 dpi. Mock is solvent control. Letters indicate the difference from Col-0. Values are means ± SD (n=6; ANOVA; p &lt; 0.01). "
NCBI gene IDs:
"PBL34: 831360
pbl34: 831360"
Figure legend: " B. LORE interacts with HopAO1 by Co-IP assays. 35S promoter-driven LORE-T7 and HopAO1-FLAG were transiently co-expressed in N. benthamiana by agroinfiltration. Plant leaves were immunoprecipated with anti-FLAG beads and immunoblotted by anti-FLAG. Coimmunoprecipitated proteins were immunoblotted by anti-T7. The experiment was repeated three times with similar results. "
NCBI gene IDs:
"35S: "
Figure legend: "  E. HopAO1 suppresses 3-OH-C10:0-triggered ROS burst. HopAO1 was ectopically expressed in Col-0 driven by estradiol-inducible promoter. 100μM estradiol was sprayed on Est:HopAO1 plants, and the leaf disc was used for ROS burst assays 12 hrs later. Values are means ± SD (n=6 biological replicates). "
NCBI gene IDs:
"HopAO1: "
Figure legend: " F. HopAO1 inhibits 3-OH-C10:0-induced disease resistance. The Est:HopAO1 plants were pretreated with 1 μM 3-OH-C10:0 (C10) and100 μM estradiol, and then the plants were inoculated with 5×104 cfu/ml Pst DC3000 24 hrs later. The bacterial titer was analyzed at 3 dpi. Values are means ± SD (n=6; ANOVA; p &lt; 0.01 "
NCBI gene IDs:
"HopAO1: "
Figure legend: "(a) RT-PCR analysis of LC3, IL-8, CHOP and GADD34 in U2OS cells subjected to glutamine deprivation (−Q) or exposed to TPG (1 μM). ("
NCBI gene IDs:
"IL-8: 3576
CHOP: 1649
LC3: 440738///81631///84557
GADD34: 23645"
Figure legend: "d) U2OS mCherry-GFP-LC3 cells were cultured in the presence (+Q) or absence (−Q) of glutamine for 18 h. Red vesicles denote autolysosomes, whereas yellow vesicles represent autophagosomes. Bars indicate numbers of yellow vesicles (autophagosomes) or red vesicles (autolysosomes) per cell±s.d. (e) Images of U2OS mCherry-GFP-LC3 cells cultured for 18 h in the presence (+Q) or absence (−Q) of glutamine. Scale bar, 10 μm."
NCBI gene IDs:
"LC3: 440738///81631///84557"
Figure legend: "(a) Immunoblot of autophagy-related proteins in U2OS cells expressing a control TALEN (U2OS-TAL2-C1) or two independent subcloned TALEN cell lines targeting ATG7 (U2OS-TAL2-A7, U2OS-TAL2-A8). p62 long and short refer to either long or short exposure times during film development."
NCBI gene IDs:
"ATG7: 10533"
Figure legend: "(b) U2OS cells expressing an ATG7-TALEN (TAL2-ATG7) or control TALEN (TAL2-CTL) were subjected to glutamine starvation for 6 days followed by 6 days recovery in glutamine-replete medium. Cells were then stained with SRB and absorbance was read at 540 nM. Graph shows average results from five clonal sub-lines for each condition. The statistical significance (P value) was determined by a two-tailed, paired Student's t-test. P0.05. (c) Representative images of SRB-stained clones in b"
NCBI gene IDs:
"ATG7: 10533"
Figure legend: "d) IL-8 enzyme-linked immunosorbent assay (ELISA) of U2OS-TAL2-CTL-C1 (control) and U2OS-TAL2-ATG7-A8 (ATG7 knockout) following 24 h glutamine deprivation."
NCBI gene IDs:
"ATG7: 10533"
Figure legend: "(d) RT-PCR analysis of IL-8 mRNA expression in U2OS cells after 24 h glutamine deprivation with or without CCI-779 or WYE-125132. Error bars in all figures represent s.d. of three biological replicates. OD, optical density."
NCBI gene IDs:
"IL-8: 3576"
Figure legend: "(b) RT-PCR analysis of IL-8 mRNA expression in U2OS cells transfected with two different siRNAs targeting JNK (JNK #1 and JNK #2) and subjected to 24 h glutamine deprivation."
NCBI gene IDs:
"IL-8: 3576
JNK: 5602///5601///5599"
Figure legend: "(d) Immunoblot analysis of JNK protein expression in JNK siRNA-treated cells."
NCBI gene IDs:
"JNK: 5602///5601///5599
JNK: P45984///P53779///P45983///5602///5601///5599"
Figure legend: "e) RT-PCR analysis of IL-8 expression in U2OS cells after 24 h glutamine deprivation with or without the JNK inhibitor SP600125 (JNKi)."
NCBI gene IDs:
"IL-8: 3576"
Figure legend: "(g) IL-8 ELISA of U2OS cells transfected with two different siRNAs targeting IRE1 (IRE-A and IRE-B) and subjected to glutamine deprivation for 24 h."
NCBI gene IDs:
"IRE1: 2081"
Figure legend: "(h) RT-PCR analysis of IL-8 mRNA expression from experiment described in g. (i) RT-PCR analysis of CHOP mRNA expression from experiment described in g. (j) RT-PCR analysis of IRE1 mRNA expression from experiment described in g."
NCBI gene IDs:
"IL-8: 3576
CHOP: 1649
IRE1: 2081"
Figure legend: "(b) RT-PCR analysis of IL-8 and CHOP mRNA expression in U2OS cells starved of glutamine for 24 h and co-treated with DMαKG."
NCBI gene IDs:
"IL-8: 3576
CHOP: 1649"
Figure legend: "(A) Primary screen anti-CD8 antibody-uptake images for control cells (top) and SFT2D2 KD cells (bottom)."
NCBI gene IDs:
"SFT2D2: 375035"
Figure legend: "(B) Cells stably expressing SFT2D2-Myc show localization of SFT2D2 to perinuclear membranes and endosomes, including VPS35-positive endosomes (arrowheads in inset). Treatment with nocodazole disperses the endosomes and even more clearly shows colocalization of SFT2D2-Myc and VPS35 (arrowheads in inset)."
NCBI gene IDs:
"perinuclear: "
Figure legend: "(D) SNARE protein staining was compared for control and SFT2D2 KD HeLa cells and quantified. The graph shows the change in cellular intensity measured for each post-Golgi SNARE investigated. Images illustrate the increased cellular intensity of VAMP3 in SFT2D2 KD cells."
NCBI gene IDs:
"SFT2D2: 375035"
Figure legend: "(E) Control and SFT2D2 KD HeLa cell lysates were separated by LDS-PAGE and blotted for the indicated SNARE proteins or actin."
NCBI gene IDs:
"SFT2D2: 375035"
Figure legend: "(A) Primary screen anti-CD8 antibody-uptake images for control cells (top) and ZDHHC5 KD cells (bottom)."
NCBI gene IDs:
"ZDHHC5: 25921"
Figure legend: "(C) Control and ZDHHC5 siRNA-treated SFT2D2-Myc cells were mixed and stained for ZDHHC5, Myc, and VPS35. KD cells are marked by an asterisk."
NCBI gene IDs:
"ZDHHC5: 25921"
Figure legend: "(D) Control and ZDHHC5 siRNA-treated HeLa cells were mixed and stained for ZDHHC5, α5-integrin, and VPS35. KD cells are indicated with an asterisk."
NCBI gene IDs:
"ZDHHC5: 25921"
Figure legend: "(E) Control (top) and ZDHHC5 KD (bottom) HeLa cells were fixed and stained for TGN46 and β1-integrin."
NCBI gene IDs:
"ZDHHC5: 25921"
Figure legend: "(A) Primary screen anti-CD8 antibody uptake images for control cells (top) and GRINA KD cells (bottom)."
NCBI gene IDs:
"GRINA: 2907"
Figure legend: "(B) Control (top) and GRINA KD (bottom) HeLa cells were stained for Golgi marker GM130 and Golgi glycoprotein-1 (GLG1)."
NCBI gene IDs:
"GRINA: 2907"
Figure legend: "(C) Cells stably expressing GFP-Rab6 were treated with GRINA siRNA (bottom) and compared to control cells (top) upon staining for TGN46 and VPS35."
NCBI gene IDs:
"GRINA: 2907"
Figure legend: "(D) Quantitation by western blotting of lysates from control and GRINA-silenced HeLa cells and GFP-Rab6 cells."
NCBI gene IDs:
"GRINA: 2907
Rab6: 84084///51560///5870"
Figure legend: "(B-E) Quantitative analysis of SFT2D2, ZDHHC5, and GRINA KD cells immunofluorescence using automated microscopy (see Experimental Procedures). (B) Representative images showing VPS35, TGN46, and CIMPR staining. Scale bar, 50 μm. (C) Quantitative analysis of CIMPR intensity at the Golgi indicates a significant increase in SFT2D2, ZDHHC5, or GRINA KD cells. (D) SFT2D2, ZDHHC5, and GRINA KD increase the Pearson's correlation coefficient for colocalization between CIMPR and retromer proteins VPS35 or SNX1 while decreasing the correlation between CIMPR and Golgi matrix protein GM130. In some cases, identical correlation coefficients were measured in the replicate experiments. (E) Quantitation of the TGN46 and GLG1 intensity in the three types of KD cells."
NCBI gene IDs:
"GRINA: 2907
SFT2D2: 375035
ZDHHC5: 25921"
Figure legend: "Human osteosarcoma U2OS cells were treated with dactinomycin (DACT) at 0.5 or 1 µM, or with mitoxantrone (MTX) between 1 and 6 µM as positive control Human osteosarcoma U2OS stably expressing CALR-GFP and H2B-RFP were treated as described above and images were acquired once per hour for 12 h (A). For one representative experiment among three, the mean ± SEM of the average area of high CALR dots (normalized to the control at each timepoint) of quadruplicates is shown (B). Values are depicted as the area under the curve ± SD of triplicates"
NCBI gene IDs:
"CALR: 
GFP: 
H2B: 
RFP: "
Figure legend: "Human osteosarcoma U2OS cells were treated with dactinomycin (DACT) at 0.5 or 1 µM, or with mitoxantrone (MTX) between 1 and 6 µM as positive control Treated U2OS cells stably expressing HMGB1-GFP and H2B-RFP images were acquired every hour for 24 h (D). For one representative experiment among three, the mean ± SEM of the green fluorescence intensity in the nucleus (normalized to the control at each timepoint) of quadruplicates is depicted (E). For each cell, the speed of nuclear release (difference of HMGB1 nuclear green fluorescence intensity between two time points) was calculated. Values are depicted as the average speed of the nuclear release ± SD of quadruplicates"
NCBI gene IDs:
"GFP: 
H2B: 
HMGB1: 
RFP: "
Figure legend: "Human osteosarcoma U2OS cells were treated with dactinomycin (DACT) at 0.5 or 1 µM, or with mitoxantrone (MTX) between 1 and 6 µM as positive control U2OS wild-type cells were treated with MTX or DACT as described above for 6 h. Then medium was refreshed and 24 h later, type I interferon response was assessed by transferring the supernatant on HT29 MX1-GFP reporter cells lines cells for additional 48 h. Human type 1α interferon (IFNα1) was also added on the cells as an additional positive control. Images were acquired by fluorescence microscopy and the number of positive cells was assessed based on the distribution of cellular green fluorescence intensity in IFNα1 versus control conditions (I). The percentage of MX1 positive cells was calculated and the mean ± SEM of five independent experiments is depicted"
NCBI gene IDs:
"GFP: 
MX1: "
Figure legend: "U2OS cells stably expressing ATF4-reporter were treated as described above for 12 h. The expression and nuclear translocation of ATF4 was assessed by fluorescence microscopy and the nuclear green fluorescence intensity was quantified. Images are shown for untreated control cells (Ctr), THAPS and DACT at 1 µM"
NCBI gene IDs:
"ATF4: 468"
Figure legend: "U2OS stably expressing ATF6-GFP were treated as described above and nuclear translocation of ATF6 was represented as the ratio of nuclear versus cytoplasmic green fluorescence intensity. Images are shown for untreated control cells (Ctr), THAPS and DACT at 1 µM"
NCBI gene IDs:
"ATF6: 
GFP: "
Figure legend: "U2OS cells stably expressing venus in frame with alternatively spliced XBP1 (sXBP1) were treated as described above for 12 h. De novo expressed venus was measured intracellular. Images are shown for untreated control cells (Ctr), THAPS and DACT at 1 µM"
NCBI gene IDs:
"venus: 
XBP1: 7494"
Figure legend: "U2OS wild-type and knock out for eIF2α kinases 1, 2, 3 and 4 cells were treated with 3 µM THAPS as a positive control for EIF2AK3-mediated eIF2α phosphorylation or with 1 µM DACT for 6 h. After fixation, cells were stained for peIF2α as described above and cytoplasmic intensity was quantified. Images are shown for untreated control cells (Ctr), THAPS and DACT at 1 µM"
NCBI gene IDs:
"eIF2α kinases 1: 27102"
Figure legend: "U2OS CALR-RFP were injected subcutaneously (s.c) in the flank of nu/nu mice. Tumors were further injected with PBS (Ctr) (n=5), 0.5 mg/kg tunicamycin (TM) for 6 h (n=3) or 0.5 mg/kg DACT for 6 h (n=4) or 24 h (n=3). Tumor slices were cut and stained with a peIF2α antibody followed by an AlexaFluor-647 secondary antibody and counterstained with Hoechst 33342. Two slices per tumor were imaged for their DAPI, RFP and Cy5 signals. Out-of-focused images were removed from the dataset leading to the analysis of the following conditions: 8 x Ctr, 5 x TM, 8 x DACT 6 h and 6 x for DACT 24 h. PeIF2α was quantified measuring Cy5 intensity in the cytoplasm (K, L) and CALR translocation by measuring the coefficient variation (CV) of the RFP signal in the cytoplasm (M, N). Representative images of peIF2α are shown for Ctr, TM and DACT at 6 h (K), whereas images of CALR are shown for Ctr and DACT at 24 h"
NCBI gene IDs:
"CALR: 
RFP: "
Figure legend: "Mice bearing WEHI 164 sarcomas were treated by systemic (intraperitoneal, i.p.) injections of DACT or PBS as a control (Ctr), and tumor were excised 9 days later for the quantitation of mRNA coding for IFNγ Two independent experiments were conducted with a total of 22 mice in the Ctr group and 23 mice in the DACT-treated group, with each data point indicating one tumor. The cycle threshold of the qRT-PCR of IFNγ was normalized to the one of the housekeeping gene peptidylpropyl isomerase A (Ppia) in each mouse and results are shown normalized with respect to controls as a dot plot depicting mean ± SD . The p-value (* p&lt;0.05) was calculated by means of a Student's t test (B)."
NCBI gene IDs:
"Ppia: 
peptidylpropyl isomerase A: 
IFNγ: 15978"
Figure legend: "A. Triple RNAscope in situ hybridization  showing Engrailed-1  (En1), Choline acetyltransferase  (ChAT) and Calbindin-1 (Calb1) expression  in the  lumbar spinal cord. En1  is expressed in  V1 interneurons, dorsal  (V1P) and ventral  (V1R) to the main ChAT-expressing motoneuron  pool. Ventral interneurons  correspond to  Renshaw cells as shown by Calb1 expression. Scale bar: 500 μm."
NCBI gene IDs:
"Calb1: 12307
Calbindin-1: 12307
Choline acetyltransferase: 12647
ChAT: 12647
En1: 13798
Engrailed-1: 13798"
Figure legend: "B.  RT-qPCR of RNA from the lumbar  enlargement at 4.5- and 16-months of age  shows stable En1  expression in WT at both ages  and a two-fold reduction of expression in heterozygous mice. Unpaired two-sided t-test. **p&lt;0.005; ***p&lt;0.0005. n=3. Values are mean ± SD."
NCBI gene IDs:
"En1: 13798"
Figure legend: "C. Triple staining EN1 IHC  (green), En1 RNAscope ISH (red) and ChAT RNAscope  (blue) demonstrating the double-staining  of En1  mRNA and protein (EN1) in the V1 interneuron  population (left panel insets, arrowheads point towards examples of double-stained V1 interneurons), and the presence of EN1  protein in large cells not expressing En1  mRNA (left panel) but expressing ChAT  (right panel insets). Scale bar: 500 µm, 30 µm for high magnification insets. D. Left: EN1  (red) detected with the LSBio antibody is localized  in ChAT-expressing neurons  (green) in the  ventral horns of the spinal cord. Right: EN1  signal is lost upon preincubation of the antibody with 1.5 M excess of recombinant hEN1. Scale bar: 50 µm."
NCBI gene IDs:
"ChAT: 12647
En1: 13798"
Figure legend: "A. Analysis of the number of En1+, Calb1+ and ChAT+ neurons  (triple RNAscope). At 4.5 months, WT and En1-Het mice  show no difference in the number of cells expressing En1, Calb1  or ChAT. Unpaired two-sided t-test. n=5-6. Data information: The analysis of neuron  number in A were performed once. Values are mean ± SD."
NCBI gene IDs:
"Calb1: 12307
ChAT: 12647
En1: 13798"
Figure legend: "D. Lumbar  level Cresyl violet staining  shows that, in contrast with small and medium size neurons  (interneurons  and γMNs, left and center panels), large size neurons  (αMNs, right panel) undergo progressive death first measured at 4.5 months. The values represent the average number of cells per  ventral horn. For the small neurons  (100-199 µm2), there was no main effect, two-way ANOVA for repeated measures for treatment group: F(1,43)=0.0017, p=0.968, ns. For the medium sized neurons  (200-299 µm2) two-way ANOVA for repeated measures for treatment group: showed no main effect F(1,43)=2.085, ns. For the large neurons (&gt;300 µm2), two-way ANOVA for repeated measures showed a significant main effect F(1,43)=59.99, p&lt;0.0001. Post-hoc comparisons were performed by unpaired two-sided t-test with equal SD comparing WT with En1-Het at each  time point (*p&lt;0.05; **p&lt;0.005; ***p&lt;0.0005; ****p&lt;0.0001). n=5 to 6. Data information: The longitudinal studies in D were performed once. Values are mean ± SD."
NCBI gene IDs:
"En1: 13798"
Figure legend: "E. Compared to WT mice, En1-Het mice  experience gradual strength loss. This loss is observed with the forepaw grip strength  (left panel), the inverted grid test (center panel and the hindlimb extensor reflex test (right panel). Strength loss is first observed between 2 and 3 months, thus before measurable αMN cell body  loss. Two-way ANOVA showed significant main effects for  grip strength  (F(1,136)=19.18, p&lt;0.0001), inverted grid test (F(1,103)=143.1, p&lt;0.0001) and extensor score  (F(1,103)=10.1, p&lt;0.0001). Comparisons were made by unpaired two-sided t-test with equal SD comparing WT with En1-Het at each  time point  (*p&lt;0.05; **p&lt;0.005; ***p&lt;0.0005; ****p&lt;0.0001). n=4 to 20. Data information: The longitudinal studies in E were performed once. Values are mean ± SD."
NCBI gene IDs:
"En1: 13798"
Figure legend: "A. NMJs  of En1-Het and WT mice  show similar numbers of AChR  clusters (left panel) and late-occurring (15.5 months) decrease in endplate  area (center panel) and percentage of perforated endplates  (right panel). The number of AChR  clusters seems to decrease between 2 and 3 months, but two-way ANOVA showed no significant genotype effect (F(1,47)=0.1291, ns). There was a significant main effect for endplate  area (F(1,47)=5.778, p=0.0202) and for the percentage of perforated endplates  (F(1,47)=13.82, p=0.0005). Post hoc analysis revealed significant genotype differences at 15.5 month of age. Unpaired two-sided t-test with equal SD comparing WT with En1-Het at each time point (**p&lt;0.005). n=4 to 8. Data information: The longitudinal study in A was performed once. Values are mean ± SD."
NCBI gene IDs:
"En1: 13798"
Figure legend: "B. Left panel illustrates the use of Alexa Fluor 488-conjugated  α-bungarotoxin (α-BTX, in green) and of neurofilament  and  synaptic vesicle glycoprotein antibodies (2H3  and SV2A, in red) to evaluate the percentage of fully occupied endplates  (&gt;80% occupancy). The right panel shows that the % of fully occupied endplates  decreases progressively in the En1-Het mouse, starting between 3 and 4.5 months of age. Scale bar: 50µm. Two-way ANOVA showed a significant main effect (F(1,47)=45.45, p&lt;0.0001). Unpaired two-sided t-test with equal SD comparing WT with En1-Het at each time point  (*p&lt;0.05; **p&lt;0.005). n=4 to 8. See details of analysis in the Methods section. Data information: The longitudinal study in B was performed once. Values are mean ± SD."
NCBI gene IDs:
"En1: 13798"
Figure legend: "A. Mice  were tested for strength at three months of age, before the onset of αMN  loss, but when strength has already decreased in En1-het mice  as measured in the forepaw grip strength,  inverted grid, and extensor reflex tests (left side of each graph). The next day, the En1-het mice  were separated into two groups. One group received buffer and the other group recombinant hEN1  (1 µg in 5 µl), injected at the L5 level. One and a half months later (4.5 months of age) En1-het mice  injected with hEN1 have recovered normal strength, in contrast with non-injected mice  or mice  injected with buffer. Unpaired two-sided t-test used at three months of age. ****p&lt;0.0001. For 4.5-month comparisons, 1-way ANOVA followed by Tukey corrected post-hoc comparisons. ***p&lt;0.0005; ****p&lt;0.0001. n=9 to 31"
NCBI gene IDs:
"En1: 13798"
Figure legend: "B. Based on the results shown in A, a new experiment was performed with a second injection 12 weeks after the first injection at 3 months of age. Again, the injection at three months of age  restored strength and in mice  receiving the second injection strength was maintained an additional ten weeks or more compared to mice  receiving a single injection. The three strength graphs demonstrate a positive effect of the second injection with values intermediate between those measured in WT mice  and En1-Het mice  with a single injection. The percentage of fully occupied endplates  and the number of αMNs  are back to wild type values in En1-Het mice  injected twice. Two-way ANOVA revealed significant main effects for grip strength (F(2,81)=25.47, p&lt;0.0001),  inverted grid (F(2,91)=51.96, p&lt;0.0001) and extensor score  (F(2,104)=30.42, p&lt;0.0001). At all times  groups were compared by Unpaired T-test with equal variances through 12 weeks. After, the groups were compared by Tukey corrected post-hoc comparisons (*p&lt;0.05; **p&lt;0.005; ***p&lt;0.0005; ****p&lt;0.0001). For the endplate  analysis and αMNs, groups were compared by 1-way ANOVA followed by Tukey corrected post hoc comparisons (*p&lt;0.05; **p&lt;0.005; ***p&lt;0.0005; ****p&lt;0.0001). n= 4 to 10."
NCBI gene IDs:
"En1: 13798"
Figure legend: "C. Intensity measurements demonstrates that the mean of SQTSM1/p62 expression  increases with age  in WT γMNs  (left) and αMNs  (right). However, comparing WT and En1-Het shows a significant difference only at 3 months and not later. Unpaired two-sided t-test. (*p&lt;0.05; **p&lt;0.005; ***p&lt;0.0005, ****p&lt;0.0001). Data information: this experiment was done once. Values are mean ± SD. Between 151 and 1215 neurons  and 3 to 5 mice were analyzed for each condition."
NCBI gene IDs:
"En1: 13798"