Journal: Science (New York, N.Y.)

Article Title: A protein network map of head and neck cancer reveals PIK3CA mutant drug sensitivity

doi: 10.1126/science.abf2911

Figure Lengend Snippet: Key Resources

Article Snippet: Gαi-DREADD (pcDNA3.1) Antibodies RSK1/2/3 antibody Cell Signaling Technology 9355 ERK1/2 Cell Signaling Technology 4695 phospho-PAK1(S199/204)/PAK2(S192/197) Cell Signaling Technology 2605 PAK1 Cell Signaling Technology 2602 PAK2 Cell Signaling Technology 2608 pERK Cell Signaling Technology 9106 FGFR3 OriGene TA801078 Daple Millipore EMD ABS515 GAPDH Cell Signaling Technology 2118 secondary goat anti-rabbit HRP Southern Biotech 4010-05 P-HER3-Y1197 Cell Signaling Technology 4561 HER3 Cell Signaling Technology 12708 goat anti-mouse HRP Southern Biotech 1010-05 anti-B-tubulin Abcam ab6276 ERK Cell Signaling Technology 9102 Deposited data Unprocessed peptide files This paper PRIDE ProteomeXchange: PXD019469 Raw data This paper PRIDE ProteomeXchange: PXD019469 Chemicals, Peptides, and Recombinant Proteins Tris G-Biosciences RC108 Acetonitrile, HPLC grade (ACN) Thermo Fisher Scientific A955-4 cOmplete protease inhibitor cocktail tablets mini, EDTA-free Roche 11846 170 001 Dithiothreitol (DTT) Sigma-Aldrich 43819 Formic acid (FA) Thermo Fisher Scientific 28905 Iodoacetamide (IAA) Acros Organic 122270250 Sequencing-grade modified trypsin Promega V5111 Benzonase Sigma E1014-25KU Trifluoroacetic acid (TFA) Thermo Fisher Scientific 28904 Urea Sigma-Aldrich U5378-1kg Fetal bovine serum (FBS) Gibco A3160502 DMEM Corning MT10013CV DMEM/F12 Corning MT10092CV Water, HPLC grade Sigma-Aldrich 270733-4 L Igepal (NP-40) Sigma-Aldrich I3021 Minimal Essential Media Corning 10-009-CV Opti-MEM Thermo Fisher Scientific 31985062 BEGM™ (Lonza) Lonza CC-3170 1% Penicillin-Streptomycin Corning MT30002Cl Paraformaldehyde, 4% solution in PBS Thermo Scientific MFCD00133991 PolyJet SignaGen SL100688 Lipofectamine 3000 ThermoFisher Scientific L3000008 hydrocortisone Sigma H6909-10ML Rapigest Waters 186001861 3x Flag Peptide Sigma F4799-4MG Anti-Flag M2 Magnetic Beads Sigma M8823-5ML Lipofectamine RNAiMAX Thermo Fisher Scientific 100014472 siFGFR3 Sigma Aldrich SIHK0780, SIHK0781, SIHK0782 native coelenterazine Biotium 10110-1 pooled siControl Dharmacon D-001810-10-20 siDaple Dharmacon L-033364-01-0005 10μM clozapine-N-oxide Cayman Chemical NC1044836 5μM native coelenterazine Biotium 10110-1 RPS6KA1 siRNA pool OriGene SR304161 non-targeting control siRNA Dharmacon D-001810-10 Triton X-100 Thermo Scientific 9002-93-1 Software and Algorithms artMS Bioconductor https://www.bioconductor.org/packages/release/bioc/html/artMS.html MSstats Bioconductor https://bioconductor.org/packages/release/bioc/html/MSstats.html Skyline MacCoss Lab https://skyline.ms/project/home/begin.view?

Techniques: Mutagenesis, Recombinant, Protease Inhibitor, Sequencing, Modification, Magnetic Beads, Software, Mass Spectrometry

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , Top enriched pathways identified by GREAT analysis of ATAC-seq and GSEA from RNA-seq in KP -O versus KP -Y primary cultures. Overlapping enriched pathways between both analyses are shown in bold ( n = 8). b , GSEA enrichment plot showing upregulation of UPR in KP -O compared with KP -Y identified in a . c , ISR pathway schematics. d , Western blot of KP -Y and KP -O in high- and low-glucose (Glc) media for the indicated time points after media renewal. e , Heatmap of ATF4 target gene expression in KP -Y and KP -O (Human Gene Set IGARASHI_ATF4_TARGETS_DN). f , Western blot of KP -Y and KP -O showing ATF4 protein levels measured after addition of 50 ng ml − 1 actinomycin D (ActD) (top), 10 µM MG-132 (middle) and combination of actinomycin D and MG-132 (bottom) at indicated time points. g , Quantification of ATF4-positive nuclei in lung tumours from KP -Young ( n = 72 tumours) and KP -Old ( n = 55 tumours). Representative ATF4 IHC images are shown. Data are median. Two-sided Kolmogorov–Smirnov test. ER, endoplasmic reticulum; FC, fold change; FDR, false discovery rate; NES, normalized enrichment score. Scale bars, 20 µm.

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques: RNA Sequencing, Western Blot, Targeted Gene Expression

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , Heatmap of core enriched genes from unfolded protein response (UPR) pathway identified by GSEA indicating the Log2FC expression in KP -Y and KP -O primary cultures ( n = 8). b , Schematic of UPR pathway. c , STRING protein–protein interaction network of UPR genes shown in ( a ), highlighting an ATF4-centered module (p < 1.1016). Main sensors of the UPR pathway IRE1α ( Ern1 ), ATF6 and ATF4 are shown. d-i , ATAC-seq tracks showing the changes in chromatin accessibility of UPR chaperone Hspa5 ( d ), UPR termination Wfs1 ( e ), ISR negative regulator Gadd34 ( f ), UPR resolution Herpud1 and Optn ( g,h ), and Atf4 ( i ). Identified peaks in KP -Y and KP -O are shown below the tracks, in green and orange respectively ( n = 3). j , Western Blot analysis of KP -Y and KP -O showing ATF4 and ATF4 upstream regulators. k , Western blot analysis of KP -Y and KP -O showing ATF4 and ATF4 downstream regulators. l , Western blot analysis of KP -Y and KP -O showing ATF4 protein levels measured at the indicated times after addition of 20 µg/ml cycloheximide (CHX). Ratio of ATF4 levels to loading control H3 represented at each time point relative to their respective level at time zero in KP -Y and KP -O ( n = 3). m , Relative mRNA expression of Atf4 at the indicated times after addition of 50 ng/ml Actinomycin D (ActD) in KP -Y and KP -O ( n = 3) primary cultures. Data are mean values ± s.e.m. Statistical significance was assessed by using Diffbind with DeSeq2 ( d-i ), Ordinary one-way ANOVA with Tukey’s multiple comparisons test ( m ).

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques: Expressing, Western Blot, Control

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , Correlation between ATF4 expression and Liver metastasis penetrance in human cell lines from DepMap ( n = 462). b-e , Relative mRNA expression of Atf4 and its targets Asns, Ddit3, Slc7a11 following twice daily treatment with 2 µM ISRIB for 72 h in KP -Y and KP -O primary cultures ( n = 3). f , KP -Y and KP -O were treated twice daily with 2 µM ISRIB for 72 h and relative viability assessed after another 72 h. g , Colony number of KP -Y ( n = 3) and KP -O ( n = 3) grown with 2 µM ISRIB for 12 days ( n = 3). h-i , Lung metastasis burden after intravenous injections of KP -Y ( n = 3) and KP -O ( n = 6) in mice and treated with either vehicle or 10 mg/kg ISRIB i.p twice daily, measured via bioluminescence (and representative ex vivo bioluminescence images of lungs, h ). Representative H&E-stained lung sections (scale bars, 2 mm, i ). H&E-quantification shown in Fig. . j , Anoikis resistance of KP -Y and KP -O ( n = 6) treated with 1 µM or 5 µM PERK inhibitor (PERKi). k , Western blot of ATF4 and ATF4 targets in sg Tom and Atf4 -KO KP -Y and KP -O. l-m , Correlation between ATF4 expression and ATF4 target gene ASNS ( l ) and SLC7A11 ( m ) in human lung adenocarcinoma (LUAD) cell lines from Dependency Map (DepMap) portal ( n = 77). n-o , IHC staining of ASNS ( n ) and SLC7A11 ( o ) in lung tumors from KP -Young and KP -Old mice 21 weeks after tumor initiation with Lenti-Cre (Scale bars, 50 µm). p-q , Anoikis resistance of KP -Y and KP -O ( n = 6) expressing either Doxycycline (dox) inducible mirE_sh Atf4 ( p ) or either sg Tom or sg Atf4 ( q ). r , Western blot analysis of ATF4 and EMT markers in KP -Y and KP -O expressing either control sg Tom or sg Atf4 . s-t , Lung metastasis burden 14 days after intravenous injection of KP -Y and KP -O expressing either sg Tom ( KP -Y sg Tom n = 4; KP -O sg Tom n = 4) or sg Atf4 ( KP -Y sg Atf4 n = 4; KP -O sg Atf4 n = 6). Scale bars, 1 mm. u , Anoikis resistance of KP -Y and KP -O expressing either control or ATF4 o/e ( n = 8). Data are mean values ± s.e.m. Simple linear regression ( a,l,m ), Ordinary one-way ANOVA with Sidak’s multiple comparisons test ( b-e ), Ordinary one-way ANOVA with Tukey’s multiple comparisons test ( g,h, j,p,q,t,u ) and two-sided unpaired t-test ( s ).

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques: Expressing, Ex Vivo, Staining, Western Blot, Immunohistochemistry, Control, Injection

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , Western blot of KP -Y and KP -O primary cultures treated with 2 μM ISRIB twice daily for 72 h. b , Anoikis resistance of KP -Y and KP -O treated with 2 µM ISRIB ( n = 6). c , Lung metastatic burden 7 days after intravenous injections of KP -Y ( n = 3 per group) and KP -O ( n = 6 per group). d , Western blot analysis of ATF4 and ATF4 targets in KP -Y and KP -O expressing a doxycycline (Dox)-inducible sh Atf4 . e , f , Lung metastatic burden 12 days after intravenous injections of KP -Y ( n = 3) and KP -O ( n = 6) expressing Dox-inducible sh Atf4 measured by means of bioluminescence ( e ), or H&E quantification ( KP -Y with or without Dox ( n = 3), KP -O EV ( n = 4 ), KP -O EV + Dox ( n = 3), KP -O with or without Dox ( n = 6)) ( f ). g , Western blot analysis of ATF4, ATF4 targets and EMT markers in KP -Y and KP -O with or without ATF4 overexpression (o/e). h , Top GSEA enriched pathways in KP -O versus KP -Y and KP -Y ATF4 o/e versus KP -Y. In bold, overlapping enriched pathways. i , Lung metastatic burden and representative H&E-stained lung sections following intravenous injections of KP -Y and KP -O expressing control or ATF4 o/e ( n = 5, 5, 4, 3). j , Western blot analysis of ATF4, ATF4 targets and EMT marker in A549 control and ATF4 o/e cells. k , Lung metastatic burden and representative H&E-stained lung sections following intravenous injections of A549 expressing either control or o/e ATF4 ( n = 5, 4). l , Heart metastasis incidence in mice injected with A549 control or ATF4 o/e cells ( n = 5). Data are mean values ± s.e.m. Ordinary one-way ANOVA with Tukey’s multiple comparisons test ( b , c , e , f , i ), two-sided unpaired t -test ( k ) and two-sided Fisher’s exact test ( l ). Scale bars, 1 mm ( f , i , k ).

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques: Western Blot, Expressing, Over Expression, Staining, Control, Marker, Injection

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , Schematic of [1,2- 13 C]- d -glucose tracing in glycolysis (blue circles) or the pentose phosphate pathway (PPP) (grey circles). b , c , Fractions of labelled pyruvate ( b ) and lactate ( c ) from [1,2- 13 C]- d -glucose in KP -Y and KP -O ( n = 6). d , Schematic of [U 13 C]- l -glutamine tracing (pink circles). e , Mass isotopomer analysis of glutamate (Glu), fumarate (Fum), citrate (Cit) and aspartate (Asp) from [U 13 C]- l -glutamine tracing in KP -Y ( n = 6) and KP -O ( n = 4). f , g , Oxygen consumption rate (OCR) ( f ) and extracellular acidification rate (ECAR) ( g ) in KP -Y ( n = 10) and KP -O ( n = 11). h , i , Mass isotopomer analysis of indicated TCA intermediates from [U 13 C]- l -glutamine tracing in KP -Y, KP -O, KP -Y sh Atf4 .2, KP -O sh Atf4 .2 ( h ) and in KP -Y; KP -Y ATF4 o/e; KP -O and KP -O ATF4 o/e ( i ) ( n = 4 per condition). j , Relative viability of KP -Y and KP -O treated with CB-839 ( n = 3) k , Heatmap of relative viability for KP -Y and KP -O treated with 0.1 μM CB-839 and the indicated compounds. All data points are relative to vehicle (Veh)-treated controls ( n = 3). l , Anoikis resistance of KP -Y; KP -Y ATF4 o/e; KP -O and KP -O ATF4 o/e treated with 0.1 μM CB-839 for 48 h ( n = 3). m , Heatmap of anoikis resistance of KP -Y and KP -O treated with 0.1 μM CB-839 and the indicated compounds. All data points are relative to vehicle-treated controls ( n = 3). n , o , Lung metastasis burden measured by means of bioluminescence ( n ) or H&E quantification ( o ) (with representative images) in mice after intravenous injections of KP -Y and KP -O and treated with CB-839 or vehicle ( n = 4, 3, 4, 5). p , q , Longitudinal tumour growth of subcutaneous tumours ( KP -Y n = 24, KP -Y + CB-839 n = 22, KP -O n = 26 and KP -O + CB-839 n = 18 tumours) ( p ) and H&E quantification of lung metastases foci ( n = 12, 11, 13 and 9 mice, respectively) ( q ). Mice were administered CB-839 once the tumours reached 100 mm 3 in size. Data are mean ± s.e.m. Two-sided multiple unpaired t -tests ( b , c , e – i , q ), two-way ANOVA ( j , p ), one-way ANOVA with Tukey’s multiple comparisons test ( l , n , o ). 5ME-THF, 5-methyltetrahydrofolate; NAC, N -acetyl- l -cysteine; NEAA, non-essential amino acids; NS, not significant. Scale bars, 1 mm.

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques:

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , Schematic depicting the fate of 13 C atoms (blue circles) from [1,2- 13 C]-D-Glucose tracing. b-c , Fractions of pyruvate ( b ) and lactate ( c ) derived from [1,2- 13 C]-D-Glucose in KP -Y and KP -O primary cultures ( n = 6). d , Mass isotopomer analysis (m1 + m2) of pyruvate, lactate and TCA cycle intermediates derived from [1,2- 13 C]-D-Glucose stable isotope tracing from KP -Y and KP -O ( n = 6). e-j , Fractions of citrate ( e ), α-Ketoglutarate ( f ), glutamate ( g ), fumarate ( h ), malate ( i ) and aspartate ( j ) isotopomers derived from [1,2- 13 C]-D-Glucose in KP -Y and KP -O ( n = 6). k , Schematic showing the metabolism of isotope-labelled [3- 13 C]-D-Glucose via pyruvate carboxylase (PC) or pyruvate dehydrogenase (PDH) pathways. Blue circles represent 13 C atoms derived from [3- 13 C]-D-Glucose. ( l-m ), Mass isotopomer analysis of pyruvate and lactate ( l ), and citrate ( m ) from 8 h [3-13 C]-D-Glucose tracing in KP -Y and KP -O ( n = 6). n , Pcx gene expression levels ( n = 8). o-r , Fractions of glutamate ( o ), fumarate ( p ), citrate ( q ) and aspartate ( r ) isotopomers derived from [U 13 C]-L-Glutamine in KP -Y ( n = 6) and KP -O primary cultures ( n = 4). s , Oxygen consumption rate (OCR) measurements using the Seahorse Cell Mito Stress Test in KP -Y and KP -O primary cultures ( n = 10 and 11). t-aa , Fractions of glutamate ( t ), fumarate ( u ), citrate ( v ) and aspartate ( w ) isotopomers from [U 13 C]-L-Glutamine in KP -Y, KP -Y sh Atf4 and KP -O, KP -O shAtf4 primary cultures ( n = 4) ( t-w ) and in control or ATF4 overexpression (o/e) in KP -Y and KP -O ( x-aa ) ( n = 4). Data are mean values ± s.e.m. Two-sided unpaired t-test ( d, l-n ).

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques: Derivative Assay, Gene Expression, Control, Over Expression

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , Schematics showing the mode of action of the inhibitors used in KP -Y and KP -O primary cultures. The enzymes and metabolites depicted in orange are ATF4 targets increased in KP -O. b-g , Relative viability assessed by CellTiter-Glo luminescence of KP -Y and KP -O after 72 h treatment with increasing doses of Erastin ( b ), ERG240 ( c ), BCATc ( d ), NCT-503 ( e ) DON ( f ), or 2-DG ( g ). All values were normalized to their respective vehicle-treated control ( n = 3). h , Colony number of KP -Y and KP -O grown in RPMI media with indicated L-Glutamine (Gln) concentrations for 6 days ( n = 6). i-j , Relative viability assessed by CellTiter-Glo luminescence of KP -Y and KP -O after 72 h treatment with increasing doses of BPTES ( i ) or V-9302 ( j ). All values were normalized to their respective vehicle-treated control ( n = 3). Data are mean values ± s.e.m. Two-way ANOVA ( b-g,i,j ), one-way ANOVA with Tukey’s multiple comparisons test ( h ).

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques: Control

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , Cell viability measured by CellTiter-Glo of KP -Y and KP -O primary cultures treated for 72 h with 1 µM ISRIB followed by 0.1 µM CB-839 treatment for another 72 h. All values were normalized to their respective vehicle-treated control ( n = 3). b , Cell viability measured by CellTiter-Glo of KP -Y and KP -O (sg Tom or sg Atf4) treated with 0.0625 µM CB-839 for 72 h. All values were normalized to their respective vehicle-treated control ( n = 3). c , Relative viability assessed by CellTiter-Glo of KP- Y and KP -O expressing either control or ATF4 o/e following 0.1 µM CB-839 treatment for 72 h. All values were normalized to their respective vehicle-treated control ( n = 3). d , Anoikis resistance of KP -Y and KP -O treated with either vehicle control, 0.1 µM CB-839, 5 µM BPTES or 2 µM V9302 ( n = 6). e , Anoikis resistance of KP -Y and KP -O treated twice a day for 72 h with 2 μM ISRIB and then seeded in ULA plates with either vehicle, 0.1 µM CB-839, 2 µM ISRIB or a combination of both ( n = 3). f , Correlation analysis between ATF4 expression and CB-839 sensitivity in human lung adenocarcinoma (LUAD) cell lines ( n = 45) from Dependency Map (DepMap) portal. Primary tumor cell lines are depicted with grey squares, metastatic cell lines with pink circles. g-i , KP -Y and KP -O subcutaneously injected in mice. Mice were administered 200 mg/kg CB-839 p.o twice/day every other day for the duration of the experiment once the tumors reached 100 mm 3 in size. Complementary results presented in Fig. and Fig. . g , Endpoint tumor weight after subcutaneous injections of KP -Y and KP -O. h , Lung metastasis burden. i , Left, H&E- stained lung sections showing metastatic foci in the lungs (black and red arrows). Right, close-up of the area identified by the red arrow in the left panel (Scale bars, 1 mm and 100 µm). Data are mean values ± s.e.m. Ordinary one-way ANOVA with Tukey’s multiple comparisons test (a,b,d,e,g,h ), two-way ANOVA ( c ) and simple linear regression ( f ).

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques: Control, Expressing, Injection, Staining

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , Flow chart of patient selection for the study. b , Distribution of age at diagnosis for patients harbouring a KRAS WT (light grey) or KRAS MUT (blue) alteration. c , Incidence of KRAS mutations frequencies in young (60 years or younger, n = 124) and older (65–75 years, n = 476) patients at diagnosis in the western Sweden cohort. d , Proportion of young (60 years or younger, green circles) or older (65–75 years, orange circles) patients from the western Sweden cohort diagnosed with stage IIIb–IV ( n = 359) NSCLC, harbouring an alteration in KRAS ( KRAS MUT ) or not ( KRAS WT ). e , Correlation analysis showing negative relation between tumour size and age at diagnosis in patients with stage IIIb–IV KRAS MUT ( n = 117) from the western Sweden cohort. f , g , Overall survival of patients with stage I–II ( n = 409) ( f ) and patients with stage III–IV ( n = 105) ( g ) LUAD from human protein atlas (TCGA dataset) stratified by high versus low ATF4 expression level. h , Correlation between percentage of ATF4-positive nuclei and age at diagnosis in patients with KRAS MUT LUAD from the Swedish NSCLC-TMA cohort ( n = 78), with shaded areas indicating 95% confidence intervals. i , ATF4 staining intensity (with representative images) in KRAS MUT LUAD tumours from young (60 years or younger, n = 26) and older patients (65 years or above, n = 47) from the Swedish NSCLC-TMA cohort. j , Survival of patients with KRAS MUT LUAD from the Swedish NSCLC-TMA cohort stratified by ATF4 staining low ( n = 35) or high ( n = 43). Two-sided Fisher’s exact test ( c ), two-sided Chi-square test ( d , i ), Simple linear regression ( e ), log-rank test ( f , g , j ) and Pearson correlation ( h ). CT, computed tomography. Scale bars, 200 μm ( i , top) and 100 μm ( i , bottom).

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques: Selection, Biomarker Discovery, Western Blot, Expressing, Staining, Computed Tomography

Journal: Nature

Article Title: Ageing promotes metastasis via activation of the integrated stress response

doi: 10.1038/s41586-026-10216-0

Figure Lengend Snippet: a , KRAS mutations frequencies in young (≤ 60 years) and old (65–75 years) patients at diagnosis in the MSK-IMPACT dataset (≤ 60 n = 145; 65–75 n = 290) (Lengel et al. ) b , Frequency of young (≤ 60 years, green circles) or old (65–75 years, gold circles) patients from the western Sweden cohort diagnosed with Stage I-IIIa, harboring an alteration in KRAS ( KRAS MUT , n = 87) or not ( KRAS WT , n = 154). c , Frequency of young (≤ 60 years, green circles) or old (65–75 years, gold circles) patients from the western Sweden cohort diagnosed with Stage I-IIIa, harboring an alteration in EGFR ( n = 83), KRAS MUT ( n = 228), all other known drivers ( BRAF, ALK, ROS1, RET ; n = 40) or unknown driver mutation ( n = 249). d-h , Correlation analyses between tumor size and age at diagnosis from the western Sweden cohort in KRAS MUT patients Stage I-IIIa ( n = 55) ( d ), KRAS WT patients (Stage I-IIIa, n = 104; Stage IIIb-IV n = 188) ( e ), EGFR MUT patients (Stage I-IIIa n = 23; Stage IIIb-IV, n = 45) ( f ), patients with unknown driver mutations (Stage I-IIIa n = 3; Stage IIIb-IV, n = 9) ( g ), patients with ALK alterations (Stage I-IIIa, n = 75; Stage IIIb-IV n = 119) ( h ). i , Positive correlation between doubling time (hours) of lung cancer cell lines and age (years) from EXPASY dataset ( n = 65). j , Correlation of Cdh1 mRNA expression and age in non-small cell lung cancer (NSCLC) cell lines from c-bioportal database ( n = 92). k , Correlation of Zeb2 mRNA expression and age at diagnosis in LUAD patients from TCGA database ( n = 491). l-m , Survival of KRAS MUT LUAD patients stratified by low or high ATF4 staining score, for young ( l ) and old ( m ) patients from Swedish NSCLC-TMA cohort. Two-sided Fisher’s exact test ( a ), two-sided Chi-square test ( b,c ), simple linear regression ( d-h, j, k ), two-tailed Spearman correlation ( i ) and Log-rank test ( l,m ).

Article Snippet: For doxycycline-induced Atf4 silencing, 1 mg ml −1 of doxycycline (no. HY-N0565B, MedChemExpress) was administered in the drinking water supplemented with 5% sucrose.

Techniques: Biomarker Discovery, Western Blot, Mutagenesis, Expressing, Staining, Two Tailed Test