EMBO/SourceData huggingface dataset:
Figure legend: "Four-week-old AG6 mice  were infected with DENV (2,500 p.f.u.) by footpad  inoculation. Blood  samples were collected for DENV viremia  detection from Day 1 to Day 7. The  viral loads from AG6 mice  infected with DENV2-Asian I  (E), were measured by qRT-PCR and were normalized against mouse GAPDH. Data information: n = 6 biological replicates. data are presented as mean ± SEM. The data were analyzed statistically using the Mann-Whitney test The P value represents a comparison between the mutants and corresponding WT within genotypes. *p &lt; 0.05, **p &lt; 0.01, ***p &lt; 0.001, n.s., not significant (p &gt; 0.05)."
NCBI gene IDs:
"GAPDH: 14433"
Figure legend: "Four-week-old AG6 mice  were infected with DENV (2,500 p.f.u.) by footpad  inoculation. Blood  samples were collected for DENV viremia   detection from Day 1 to Day 7. The  viral loads from AG6 mice  infected with DENV2-Asian American (F), DENV3-I (G) were measured by  qRT-PCR  and were normalized against mouse GAPDH. Data information: , n = 6 biological replicates. In (F), n = 5 biological replicates. data are presented as mean ± SEM. The data were analyzed statistically using the Mann-Whitney test The P value represents a comparison between the mutants and corresponding WT within genotypes. *p &lt; 0.05, **p &lt; 0.01, ***p &lt; 0.001, n.s., not significant (p &gt; 0.05)."
NCBI gene IDs:
"GAPDH: 14433"
Figure legend: "Four-week-old AG6 mice  were infected with DENV (2,500 p.f.u.) by footpad  inoculation. Blood  samples were collected for DENV viremia   detection from Day 1 to Day 7. The viral loads from AG6 mice  infected with DENV4-I  (H) were measured by qRT-PCR  and were normalized against mouse GAPDH. Data information: n = 6 biological replicates. data are presented as mean ± SEM. The data were analyzed statistically using the Mann-Whitney test The P value represents a comparison between the mutants and corresponding WT within genotypes. *p &lt; 0.05, **p &lt; 0.01, ***p &lt; 0.001, n.s., not significant (p &gt; 0.05)."
NCBI gene IDs:
"GAPDH: 14433"
Figure legend: "I, J Infection  of DENV2-Asian I  mutants on  hu-moDC  (I) and  hu-moMØ  (J). The cells were infected with 0.01 MOI of the indicated viruses. The infected cells were detected at 48 h postinfection by  qRT-PCR, and DENV viral loads were normalized against human GAPDH. Data information: n = 6 biological replicates. data are presented as mean ± SEM. The data were analyzed statistically using the unpaired t-test (I, J). The P value represents a comparison between the mutants and corresponding WT within genotypes. *p &lt; 0.05, **p &lt; 0.01, ***p &lt; 0.001, n.s., not significant (p &gt; 0.05)."
NCBI gene IDs:
"GAPDH: 2597"
Figure legend: "B The 293T  cells were transfected with a pcDNA3.1 recombinant plasmid with a human DC-SIGN gene. The cells transfected by an empty pcDNA3.1 plasmid served as negative controls. Thirty-six hours after transfection, the cells were infected with DENV2  (0.01 MOI). The DENV2 viral loads were assessed at 48 h postinfection via qRT-PCR and were normalized against human GAPDH. Data information: In (B), n = 6 biological replicates. In (B, the data are presented as means ± SEM and were analyzed statistically using an unpaired t-test. The P value represents a comparison between the mutants and the 16681 WT strain. *p &lt; 0.05, **p &lt; 0.01, ***p &lt; 0.001, ****p &lt; 0.0001, n.s., not significant (p &gt; 0.05). "
NCBI gene IDs:
"DC-SIGN: 30835
GAPDH: 2597"
Figure legend: "A RNF123 suppresses the SeV- and EMCV-induced but not the cGAS and STING-mediated activation of the IFN-βpromoter. HEK293T cells (3×105) were transfected with the IFN-β reporter plasmid (100 ng), the pRL-TK Renilla luciferase plasmid (100 ng), the expression plasmid for cGAS (50 ng) and STING (50 ng) or empty vector (100 ng), and 0, 20, 50, or 100 ng (wedges) of RNF123 for 12 h, and then were infected with SeV or EMCV or left uninfected. Luciferase assays were performed after 18 h and results are presented relative to the luciferase activity in control cells."
NCBI gene IDs:
"luciferase: 
Renilla luciferase: 
IFN-β: 3456
cGAS: 115004
RNF123: 63891
STING: 340061"
Figure legend: "B RNF123 inhibits the SeV-, but not the cGAS and STING-induced activation of the NF-κB promoter. HEK293T cells (3×105) were transfected with the NF-κB reporter plasmid (100 ng), the pRL-TK Renilla luciferase plasmid (100 ng), the expression plasmids for cGAS (50 ng) and STING (50 ng), or empty vector (100 ng), and 0, 20, 50 or 100 ng (wedges) of RNF123 for 12 h, and were then infected for 24 h with SeV or left uninfected, followed by analysis as described in A."
NCBI gene IDs:
"Renilla luciferase: 
cGAS: 115004
NF-κB: 4791///4790
RNF123: 63891
STING: 340061"
Figure legend: "C, D RNF123 inhibits the SeV-induced, but not the cGAS and STING-mediated RANTES and IFN-β transcription. The transfection and treatment were as in A. Total RNA was isolated and RT-PCR (C) was performed using the indicated primers."
NCBI gene IDs:
"RANTES: 6352
IFN-β: 3456
cGAS: 115004
RNF123: 63891
STING: 340061"
Figure legend: "C, D RNF123 inhibits the SeV-induced, but not the cGAS and STING-mediated RANTES and IFN-β transcription. The transfection and treatment were as in A. Total RNA was isolated and Q-PCR (D) was performed using the indicated primers."
NCBI gene IDs:
"IFN-β: 3456
cGAS: 115004
RNF123: 63891
STING: 340061"
Figure legend: "E RNF123 inhibits the SeV-induced, but not the cGAS and STING-mediated IFN-β secretion. The transfection and treatment were as in A. Bioassays for type I interferon in the supernatants of HEK293T cells were performed and the results are presented relative to untreated control cells."
NCBI gene IDs:
"cGAS: 115004
RNF123: 63891
STING: 340061"
Figure legend: "F Overexpression of RNF123 enhances NDV-eGFP replication. HEK293T cells (1×105) were transfected with the expression plasmids for Flag-RNF123 or empty vector (2 µg). At 24 h after transfection, cells were infected with NDV-eGFP. At 30 h after infection, viral replication was determined by GFP expression visualized by fluorescence microscopy and analyzed by western blot."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "G RNF123 enhances EMCV replication. HeLa cells (1×105) were transfected with the expression plasmids for Flag-RNF123 or empty vector (2 µg). At 24 h after transfection, cells were infected with EMCV or given DMEM with no virus. At 18 h after infection, viral replication was determined by VP1 expression, which was analyzed by western blot."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "A HEK293T cells with stable knockdown of RNF123 were generated using shRNA. Western blot was performed with anti-RNF123, anti-VISA (as a control), and anti-GAPDH antibodies after infection with five lentiviruses encoding a shRNA control (GFPsh) and four shRNAs targeting RNF123 (RNF123 sh #1, #2, #3, and #4) in HEK293T cells."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "B, C Knockdown of RNF123 facilitated the activation of IFN-β and NF-κB promoters induced by SeV and EMCV but not by cGAS and STING. Stable HEK293T cells with GFPsh or RNF123 shRNAs (2×105) were transfected with the indicated reporter plasmid (100 ng), pRL-TK Renilla luciferase plasmid (100 ng), cGAS (50 ng), and STING (50 ng), or empty vector (100 ng). At 12 h after transfection, they were infected for 12 h with SeV or EMCV or left uninfected, followed by analysis as described in the legend of Fig 1A."
NCBI gene IDs:
"Renilla luciferase: 
IFN-β: 3456
cGAS: 115004
NF-κB: 4791///4790
RNF123: 63891
STING: 340061"
Figure legend: "D, E Knockdown of RNF123 enhances the IFN-β and RANTES transcription induced by SeV and EMCV, but not that by cGAS and the STING. HEK293T cells were transfected with cGAS (1 μg) and STING (1 μg) or empty vector for 12 h, and then infected with SeV or EMCV for 12 h or left uninfected. Total RNA was isolated and RT-PCR (D) were performed using the indicated primers."
NCBI gene IDs:
"RANTES: 6352
IFN-β: 3456
cGAS: 115004
RNF123: 63891
STING: 340061"
Figure legend: "D, E Knockdown of RNF123 enhances the IFN-β and RANTES transcription induced by SeV and EMCV, but not that by cGAS and the STING. HEK293T cells were transfected with cGAS (1 μg) and STING (1 μg) or empty vector for 12 h, and then infected with SeV or EMCV for 12 h or left uninfected. Total RNA was isolated and Q-PCR (E) were performed using the indicated primers."
NCBI gene IDs:
"IFN-β: 3456
cGAS: 115004
RNF123: 63891
STING: 340061"
Figure legend: "F, G Time-course of potentiation of SeV-induced IFN-β transcription (F) by knockdown of RNF123. HEK293T cells were infected with SeV for 6, 12, 18, or 24 h, total RNA was isolated, and RT-PCR was performed using the indicated primers (F)."
NCBI gene IDs:
"IFN-β: 3456
RNF123: 63891"
Figure legend: "F, G Time-course of potentiation of SeV-induced IFN-β secretion (G) by knockdown of RNF123. HEK293T cells were infected with SeV for 6, 12, 18, or 24 h, total RNA was isolated. Bioassays of type-I interferon in the supernatants of HEK293T cells were performed, and the results are presented relative to untreated control cells (G)."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "H HeLa cells with stable knockdown of RNF123 were generated by shRNA. Western blot was performed as described in Fig 2A."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "I, J Knockdown of RNF123 enhances SeV- and EMCV- induced IFN-β expression in HeLa cells. HeLa cells were infected with SeV or EMCV for 12 h, total RNA was isolated, and RT-PCR (I) was performed using the indicated primers."
NCBI gene IDs:
"IFN-β: 3456
RNF123: 63891"
Figure legend: "I, J Knockdown of RNF123 enhances SeV- and EMCV- induced IFN-β expression in HeLa cells. HeLa cells were infected with SeV or EMCV for 12 h, total RNA was isolated, and Q-PCR (J) was performed using the indicated primers."
NCBI gene IDs:
"IFN-β: 3456
RNF123: 63891"
Figure legend: "K Knockdown of RNF123 suppresses NDV-eGFP replication. HEK293T cells (2×105) were infected with NDV-eGFP. At 30 h after infection, viral replication was determined by GFP expression visualized by fluorescence microscopy and analyzed by western blot with GFP antibody."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "L Knockdown of RNF123 inhibits EMCV replication. HeLa cells (2×105) were infected with EMCV for 0, 12, or 18 h."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "A, B RNF123 inhibits the activation of the IFN-β (A) promoters induced by the overexpression of RIG-I, RIG-I CARD, MDA5 CARD, and VISA, but has no effect on the TBK1- and IRF3-5D-induced activation of the IFN-β promoter (A) . HEK293T cells (2×105) were transfected with the indicated plasmids (100 ng), IFN-β promoter luciferase plasmid (100 ng), and pRL-TK Renilla luciferase plasmid (100 ng), together with an empty vector and RNF123 construct (100 ng). Luciferase assays were performed 24 h after transfection."
NCBI gene IDs:
"Renilla luciferase: 
RIG-I: 23586
MDA5: 64135
IFN-β: 3456
IRF3: 3661
VISA: 57506
RNF123: 63891
TBK1: 29110"
Figure legend: "A, B RNF123 inhibits the activation of the NF-κB (B) promoters induced by the overexpression of RIG-I, RIG-I CARD, MDA5 CARD, and VISA, but has no effect on the TAB2- and IKKβ-induced activation of the NF-κB promoter (B). HEK293T cells (2×105) were transfected with the indicated plasmids (100 ng), NF-κB promoter luciferase plasmid (100 ng), and pRL-TK Renilla luciferase plasmid (100 ng), together with an empty vector and RNF123 construct (100 ng). Luciferase assays were performed 24 h after transfection."
NCBI gene IDs:
"luciferase: 
Renilla luciferase: 
RIG-I: 23586
MDA5: 64135
IKKβ: 3551
NF-κB: 4791///4790
RNF123: 63891
TAB2: 23118"
Figure legend: "C Knockdown of RNF123 potentiates the activation of the IFN-β promoter induced by RIG-I, RIG-I CARD, MDA5, and MDA5 CARD, but not that by VISA, TBK1, and cGAS/STING. 293T-GFPsh, shRNF123 #1, #3, and #4 stable cell pools (2×105) were transfected with the indicated plasmids. Luciferase assays were performed 24 h after transfection."
NCBI gene IDs:
"RIG-I: 23586
MDA5: 64135
IFN-β: 3456
VISA: 57506
cGAS: 115004
RNF123: 63891
TBK1: 29110
STING: 340061"
Figure legend: "D RNF123 interacted with RIG-I and MDA5, but not VISA, TBK1, and cGAS. HEK293T cells (1×106) were transfected with Flag-RNF123 (F-RNF123) together with HA-RIG-I, HA-MDA5, HA-VISA, HA-TBK1, or HA-cGAS plasmids (8 μg each). Cell lysates were immunoprecipitated (IP) with anti-HA (αHA). The immunoprecipitates were analyzed by western blotting (immunoblotting [IB]) with anti-Flag (αF) or anti-HA antibody (αHA). Whole-cell lysates (WCL) were analyzed by western blotting with anti-HA and anti-Flag antibodies to determine the expression levels of transfected plasmids."
NCBI gene IDs:
"RIG-I: 23586
MDA5: 64135
VISA: 57506
cGAS: 115004
TBK1: 29110"
Figure legend: "B The SPRY and coiled-coil domains are both indispensable for inhibiting IFN-β promoter activation. HEK293T cells (3×105) were transfected with the IFN-β reporter plasmid (100 ng), the pRL-TK Renilla luciferase plasmid (100 ng), the expression plasmid for RNF123 and its mutants (100 ng), and the empty vector or RIG-I CARD/MDA5 CARD (100 ng). Reporter assays were performed 24 h after transfection."
NCBI gene IDs:
"Renilla luciferase: 
RIG-I: 23586
MDA5: 64135
IFN-β: 3456
RNF123: 63891"
Figure legend: "C The SPRY domain and two coiled-coil domains of RNF123 are both required for interaction with RIG-I CARD. HEK293T cells (1×106) were transfected with HA-RIG-I CARD together with F-RNF123 or its mutants (8 μg each). Cell lysates were immunoprecipitated (IP) with anti-Flag (α-F). The immunoprecipitates were analyzed by western blotting (immunoblotting [IB]) with anti-HA (α-HA) or anti-Flag (α-F). Whole-cell lysates (WCL) were analyzed by western blotting with anti-HA and anti-Flag antibodies."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "D KPC2 has no effect on the activation of the IFN-β reporter induced by RIG-I CARD and MDA5 CARD. HEK293T cells (3×105) were transfected with the IFN-β reporter plasmid (100 ng), the pRL-TK Renilla luciferase plasmid (100 ng), and the indicated expression plasmids. Reporter assays were performed 24 h after transfection."
NCBI gene IDs:
"Renilla luciferase: 
RIG-I: 23586
MDA5: 64135
IFN-β: 3456
KPC2: 10422"
Figure legend: "E Effect of shKPC2 plasmids on endogenous KPC2. RT-PCR was performed with the indicated primers after HEK293T cells were infected with six lentiviruses encoding an shRNA control (GFPsh) and five shRNAs targeting kpc2 (Kpc2 #1-#5)."
NCBI gene IDs:
"KPC2: 10422
kpc2: 10422
Kpc2: 10422"
Figure legend: "F KPC2 is not required for the inhibitory action of RNF123. HEK293T cells (2×105) with GFPsh or KPC2 sh (#1, #2, or #4) were transfected with the IFN-β reporter plasmid (100 ng) and the pRL-TK Renilla luciferase plasmid (100 ng) with the empty vector or RNF123 (100 ng). Twenty-four hours after transfection, cells were infected with SeV or left uninfected. Reporter assays were performed 18 h after infection."
NCBI gene IDs:
"Renilla luciferase: 
IFN-β: 3456
RNF123: 63891
KPC2: 10422"
Figure legend: "G Knockdown of KPC2 had no effect on SeV-induced IFN-β expression. HEK293T cells with GFPsh or KPC2 sh #4 were infected with SeV for the indicated times, total RNA was isolated, and RT-PCR experiments were performed."
NCBI gene IDs:
"IFN-β: 3456
KPC2: 10422"
Figure legend: "A, B RNF123 interacts with RIG-I/MDA5 CARD. HEK293T cells (1×106) were transfected with expression plasmids for Flag-RNF123 together with HA-RIG-I or its truncations (8 μg each). Co-immunoprecipitation and whole-cell lysates were analyzed as described in the legend of Fig. 3D."
NCBI gene IDs:
"RIG-I: 23586
MDA5: 64135"
Figure legend: "C, D RNF123 competes with VISA for RIG-I/MDA5 CARD binding. HEK293T cells (1×106) were transfected with expression plasmids for HA-RIG-I CARD/MDA5 CARD alone or together with Flag-VISA or with Flag-VISA and Flag-RNF123 (8 μg each). Co-immunoprecipitation and whole-cell lysates were analyzed as described in Fig. 3D."
NCBI gene IDs:
"RIG-I: 23586
MDA5: 64135
VISA: 57506
RNF123: 63891"
Figure legend: "E RNF123 does not compete with REUL for RIG-I CARD binding. The methods of transfection and co-immunoprecipitation were as described in Fig. 5C,D."
NCBI gene IDs:
"RNF123: 63891
REUL: 84282"
Figure legend: "F Knockdown of RNF123 facilitates the endogenous interaction between RIG-I and VISA in 293T cells. 293T cells stably infected with SeV were treated for the indicated times. Cell lysates were immunoprecipitated with anti-VISA antibody. The immunoprecipitates and lysates were analyzed by western blotting with the indicated antibody."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "G Knockdown of RNF123 facilitates the endogenous interaction between MDA5 and VISA in HeLa cells. HeLa cells were stably infected with EMCV for the indicated times. Co-immunoprecipitation and whole-cell lysates were analyzed as described in (F)."
NCBI gene IDs:
"RNF123: 63891"
Figure legend: "C) 8 weeks old C57BL/6N male mice were fasted (food removed at ZT 12) or re-fed (fasted from ZT4 to ZT12 for synchronization; food re-introduced at ZT 12). Tissue samples were collected at ZT 12, 15, 18 and 21 (n=4). Liver lysates analyzed by immunoblotting using anti-Prox1 and anti-P_S6K(Thr389) antibodies; β actin was detected as an input control. The quantification of SUMOylated Prox1 (%) and total Prox1 protein expression as well as Prox1 mRNA levels analyzed by qPCR are shown. qPCR data is presented as relative fold change normalized to the housekeeping gene TBP. Data information: Every dot represents one individual mouse. Data: mean ±SEM. Significance was determined by one-way ANOVA with Dunnett´s multiple comparison test relative to samples collected at ZT 12. *** P≤0.001."
NCBI gene IDs:
"Prox1: 19130
TBP: 21374"
Figure legend: "B) HepG2 cells overexpressing HA-tagged mouse wild-type Prox1 (wt), Prox1 K556R mutant or Prox1 E558A mutant. Cell lysates analyzed by immunoblotting using anti-HA antibodies; tubulin was detected as an input control."
NCBI gene IDs:
"HA: 
Prox1: 19130"
Figure legend: "B) 8 weeks old Prox1K556R K.I. (f/f) male mice were injected with a control AAV (Ctrl_AAV), with an AAV to overexpress Cre recombinase (Cre_AAV) or with phosphate-buffered saline (PBS) (n=4). Liver tissue samples were collected 3 weeks later from mice fed ad libitum. Liver lysates were analyzed by immunoblotting using anti-Prox1 and anti-Cre antibodies; Vcp was detected as an input control. The quantification of SUMOylated Prox1 and total Prox1 protein expression as well as Prox1 mRNA levels analyzed by qPCR are shown. qPCR data presented as relative fold change normalized to the housekeeping gene TBP. Data information: (B) Every dot represents one individual mouse. Data: mean ±SEM. Significance was determined by one-way ANOVA with Tukey´s multiple comparison test between groups. *** P≤0.001, **** P≤0.0001."
NCBI gene IDs:
"Cre: 2777477
Prox1: 19130
TBP: 21374"
Figure legend: "A) 8 weeks old Prox1K556R K.I. (f/f) male mice were injected with a control AAV (Ctrl_AAV) or with an AAV to overexpress Cre recombinase (Cre_AAV) and placed on a high-fat (45%) high-fructose (20% w/v) diet for 18 weeks. Liver samples were collected between ZT 16 and 17 in the fasted (8 h) and re-fed (fasted 8 h and re-fed 4-5 h) states (n=7-8). Liver lysates were analyzed by immunoblotting using anti-Prox1 and anti-Cre antibodies; β actin was detected as an input control. The quantification of SUMOylated Prox1 and total Prox1 protein expression are shown. Data information Every dot represents one individual mouse. Data: mean ±SEM. Significance was determined by two-way ANOVA with Sidak´s multiple comparison test between different groups and conditions. * P≤0.05, ** P≤0.01, **** P≤0.0001."
NCBI gene IDs:
"Cre: 2777477
Prox1: 19130"
Figure legend: "C) Heatmap displaying the expression of enriched genes involved in phase I, II and III of bile acid detoxification and bile acid synthesis in the liver of obese mice expressing the Prox1 K556R mutant (Cre_AAV) in comparison to obese mice expressing wild-type Prox1 (Ctrl_AAV) in the fasted state (n=4). The occupation scores according to a published Chip- sequencing analysis on Prox1 Cyp7a1 was used as a reference gene."
NCBI gene IDs:
"Cre: 2777477
Cyp7a1: 13122
Prox1: 19130"
Figure legend: "D) Prox1/LRH-1 co-immunoprecipitation in HEK293A cells overexpressing HA-tagged Prox1 wild-type (wt) or K556R mutant (KR) and Myc-tagged LRH-1."
NCBI gene IDs:
"HA: 
Myc: 
LRH-1: 26424
Prox1: 19130"
Figure legend: "C The parental population and the NFR resistant clones were analyzed for eEF2K mRNA expression by real-time PCR relative to β-actin (mean and s.e.m. from 3 independent batches of cells are shown). P values were calculated using 1way ANOVA analysis of variance followed by Dunnett's multiple comparison tests. *P value ≤ 0.05, ** P value ≤ 0.01, *** P value ≤ 0.001."
NCBI gene IDs:
"β-actin: 
eEF2K: 29904"
Figure legend: "B eEF2K WT and KO MEF treated for 6h with indicated concentration of NFR, 200 nM rapamycin (Rapa.), 1h starvation (Starv.) or 1 mM of the AMPK activator AICAR, were analyzed by immunoblot with antibodies directed against total or phosphorylated eEF2 (Thr56)."
NCBI gene IDs:
"eEF2K: 13631"
Figure legend: "B eEF2K WT and KO MEF treated for 6h with indicated concentration of NFR, Rapamycin (Rapa), AICAR or starved for indicated time in PBS (Starv.), were analyzed by immunoblot using indicated antibodies. Tubulin is used as loading control."
NCBI gene IDs:
"eEF2K: 13631"
Figure legend: "D NFR-mediated eEF2 phosphorylation is not affected in AMPKα1α2 dKO. AMPKα1α2 WT and dKO MEF treated for 6h with indicated concentration of NFR, Rapamycin (Rapa), AICAR or starved for 1h in PBS (Starv.), were analyzed by immuoblot with indicated antibodies. * anti-total AMPKα antibody give an unspecific band with a slightly higher molecular weight in dKO cells. Tubulin is used as loading control. Each panel is representative of at least 3 independent experiments."
NCBI gene IDs:
"AMPKα1: 105787"
Figure legend: "A-B EEF2K contributes to the decreased translation observed with NFR without impacting translation in presence of TM. A. Quantification of newly synthesized proteins at 0, 2, 4 and 6h after 20 μM NFR (left panels) or 10 μg/ml TM (right panels) treatment in MEF eEF2K +/+ and eEF2K-/-. Treated cells were labeled for 15 minutes with 35S-methionine and visualized by SDS PAGE and subsequent autoradiography. Autoradiographies from four different experiments (see also source data) were quantified and results show percentage of translation compared to untreated cells. The mean and s.e.m of four independent metabolic labeling experiments are shown. *P value ≤ 0.05, ** P value ≤ 0.01, *** P value ≤ 0.001 obtained using 2way ANOVA analysis of variance (in red) followed by Bonferroni posttest (in black). B. eEF2K+/+, eEF2K-/-, eIF2αWT and eIF2αS51A MEFs were treated for 6h with indicated doses of NFR, 200 nM rapamycin, or 10 μg/ml TM, or for 30 min with 10 μg/ml CHX. 35S-methionine incorporation was measured by liquid scintillation counting. Data are shown as the percentage of translation compared to untreated cells. *P value ≤ 0.05, ** P value ≤ 0.01, *** P value ≤ 0.001 obtained using 2way ANOVA analysis of variance followed by Bonferroni posttest."
NCBI gene IDs:
"eEF2K: 13631
EEF2K: 13631
eIF2α: 13665"
Figure legend: "C Representative polysome profiles of eEF2K WT and KO MEF treated 6h with NFR or TM. Area under curve for Sub-polysomes (S) and Polysomes (P) used to calculate the P/S ratio were indicated (see also raw data provided). Bar graph represents ratio normalized to untreated cells. OD254 nm is optical density at 254 nm. Data showed mean ±s.e.m. of P/S ratio calculated from 3 independent experiments. P values were calculated using 1way ANOVA analysis of variance followed by Bonferroni posttest; *P value ≤ 0.05, ** P value ≤ 0.01."
NCBI gene IDs:
"eEF2K: 13631"