Journal: Nature Metabolism

Article Title: Tissue-specific fibroblast lipid cues impose the rate of epithelial cancer invasion

doi: 10.1038/s42255-026-01514-y

Figure Lengend Snippet: a , SM metabolic pathway and inhibitors targeting enzymes. b – d , Spheroid invasion of oSCC with control media (grey), 5 µM SM (blue) or 5 µM SM and 20 µM imipramine (yellow) ( b ), oral FB secretomes (grey), with 20 µM imipramine (blue) or 20 µM imipramine and 2 µM S1P (yellow) ( c ) and oral FB secretomes (grey), with 2 µM PF543 (blue) or 2 µM PF543 and 2 µM S1P ( d ) (two-sided Mann–Whitney U- test, n = 6 replicates, two cell lines, two independent experiments). e , S1P immunofluorescence quantification of FADU control (grey), dermal (yellow) and oral (blue) FB secretomes (two-sided Mann–Whitney U- test, n = 6 replicates, three independent FB secretomes). f , Immunofluorescence images of FADU + dermal (left) or oral (right) FB secretomes; blue, Hoechst; orange, phalloidin; green, S1P; scale bar, 20 µm (representative images from n = 6 biologically independent wells). g , Spheroid invasion of oSCC cells treated with oral FB secretomes (grey) with 5 µM S1PR1 inhibitor (S1PR1i, ponesimod, blue) or 10 µM S1PR3 inhibitor (S1PR3i, TY-52156, yellow; Kruskal–Wallis, Dunn’s multiple comparisons, n = 5 replicates). h , Spheroid invasion of oSCC treated with oral FB secretomes (grey) with 20 µM STAT3 inhibitor (STAT3i, STX-0119, blue), 10 µM NF-κB inhibitor (NF-κBi; JSH-23, yellow) or 2 µM AKT inhibitor (AKTi; MK-2206, red; Kruskal–Wallis, Dunn’s multiple comparisons, n = 5 replicates). i , Western blot of STAT3, phospho-STAT3 Tyr705 , B-actin in oSCC (FADU) + dermal or oral FB secretomes, ±PF543, S1P rescue, S1PR1 inhibitor (ponesimod) or STAT3 inhibitor (STX-0119); numbers represent normalized pSTAT3 to total STAT3 fold change relative to oral FB secretomes, run on separate blots with independent loading controls under identical conditions (western blots representative of two independent experiments). j , oSCC basal respiration in oSCC cells treated with oral FB secretomes ± lipid stripping (Cleanascite, blue, 24 h; OCR, oxygen consumption rate; n = 24 replicates in two cell lines treated with three FB secretomes, two independent experiments, two-sided Mann–Whitney U- test). k , Mitochondrial activity in oSCC cell lines treated with oral FB secretomes (blue) measured by membrane potential-dependent immunofluorescence (two-sided Mann–Whitney U- test, n = 8 replicates, two independent cell lines). l , Mitochondrial abundance in oSCC cell lines treated with oral FB secretomes (blue) measured by immunofluorescence (two-sided Mann–Whitney U- test, n = 8 replicates, two independent cell lines). m , Basal respiration in oSCC cells treated with combinations of oral FB secretomes, 20 µM imipramine, 2 µM S1P or 20 µM STAT3i (STX-0119; n = 5 independent replicates, two-sided Mann–Whitney U- test). n , Subcutaneous tumour growth rate in NSG mice injected with oSCC UMSCC01 (grey), UMSCC01 + dermal FBs (oSCC + dermal FB, yellow) or UMSCC01 + oral FBs (oSCC + oral FB, blue; two-sided Mann–Whitney U- test, n = 7 per group). o , Ki67 proliferation quantification in oSCC (grey), oSCC + dermal FB (yellow) and oSCC + oral FBs (blue) tumours (two-sided Mann–Whitney U- test, n = 14, two measurements per tumour). p , Kaplan–Meier survival curve of oSCC (black) and oSCC + oral FBs (blue; n = 7 per group, two-sided Mantel–Cox test). Box plots show minimum to maximum values (error bars), the box indicates 25th and 75th percentiles, and the line denotes the median.

Article Snippet: STAT3 inhibitor STX-0119 (MedChemExpress, HY-103692-1ml), AKT inhibitor MK-2206 (MedChemExpress, HY-10358-1ml) and NF-κB inhibitor JSH-23 (MedChemExpress, HY-13982-1ml) were supplied at 10 mM in DMSO and diluted to 20 μM, 2 μM and 10 μM, respectively, in secretomes.

Techniques: Control, MANN-WHITNEY, Immunofluorescence, Western Blot, Stripping Membranes, Activity Assay, Membrane, Injection

Journal: Nature Metabolism

Article Title: Tissue-specific fibroblast lipid cues impose the rate of epithelial cancer invasion

doi: 10.1038/s42255-026-01514-y

Figure Lengend Snippet: a , oSCC spheroid invasion with oral fibroblast secretome treated with (blue) or without (grey) 5 µM ARC39 (two-sided Mann Whitney U, n = 6 replicates, two cell lines, two independent experiments). b , oSCC (FADU) spheroid invasion with oral fibroblast secretome (grey) with 5 µM (blue) and 10 µM (yellow) carmofur (Kruskal-Wallis, Dunn’s multiple comparisons, Oral FB, 10 µM: n = 8 replicates, 5 µM: n = 7). c , oSCC spheroid invasion +/- oral fibroblast secretomes control (grey), 20 µM SPHK2 inhibitor ABC294640 (blue) (two-sided Mann Whitney U, ns: non-significant, n = 5 replicates, two cell lines, two independent experiments). d , Spheroid invasion of oSCC with control media (grey) or sphingsine-1-phosphate (S1P, 1 µM yellow, 2 µM blue), (two-sided Mann Whitney U, n = 6 replicates, two cell lines, two independent experiments). e , Proliferation (relative confluence) of oSCC treated with 20 µM sphingomyelin (SM) and with 20 µM imipramine or 2 µM PF543 for 24 h (two-sided Mann Whitney U, n = 5 replicates, bars represent mean, error bars SEM). f , Replicate western blot of STAT3, phospho-STAT3Tyr705, B-actin in oral SCC (FADU) + dermal or oral fibroblast secretome, +/- PF-543, S1P rescue, S1PR1 inhibitor (ponesimod), or STAT3 inhibitor (STX-0119), run on separate blots with independent loading controls under identical conditions. g , Fold change in pSTAT3 levels relative to oral fibroblast secretome (Oral sec) in FADU western blots (n = 2 independent western blots). h , Relative expression (RE) of EMT genes VIM, FN1, and SNAI2 in oral SCC cells treated with oral fibroblast secretome (grey) and 20 µM STAT3 inhibitor (STAT3i, blue) STX-0119 (two-sided Mann Whitney U, n = 9 replicates across three independent samples). i , Oxygen consumption rate (OCR) plots of Seahorse assay of oSCC treated with fibroblast secretome ± Cleanascite (control: n = 8 biological replicates, oral FB: n = 24 replicates in two cell lines treated with three fibroblast secretomes). j , ATP production in oSCC cells treated with oral fibroblast secretomes +/- lipid stripping (Cleanascite: blue) (OCR: oxygen consumption rate, n = 24 replicates in two cell lines treated with three fibroblast secretomes, two independent experiments, two-sided Mann Whitney U). k , Basal respiration and ATP production ( l ) in oSCC (FADU) treated with sphingomyelin (SM), sphingosine-1-phosphate (S1P) or oral fibroblast secretome (24 h, two-sided Mann Whitney U, n = 5 replicates). m , Oxygen consumption rate (OCR) plots of Seahorse assay of FADU treated with sphingomyelin (SM), sphigosine-1-phosphate (S1P) or oral fibroblast secretome (n = 5 biological replicates). n , Representative immunofluorescence images of oSCC mitochondria stained with MitoView 633 (red), before and after exposure to oral fibroblast secretome, scale = 25 µm. o , Basal respiration of FADU cells treated with fibroblast secretome, Cleanascite treated secretome and rescue with addition of 2 µM S1P (Kruskal-Wallis, Dunn’s multiple comparisons, n = 5 replicates). Boxplots: minimum to maximum values (error bars), box: 25th and 75th percentiles; line: median. Bar plots display mean ± standard error of the mean.

Article Snippet: STAT3 inhibitor STX-0119 (MedChemExpress, HY-103692-1ml), AKT inhibitor MK-2206 (MedChemExpress, HY-10358-1ml) and NF-κB inhibitor JSH-23 (MedChemExpress, HY-13982-1ml) were supplied at 10 mM in DMSO and diluted to 20 μM, 2 μM and 10 μM, respectively, in secretomes.

Techniques: MANN-WHITNEY, Control, Western Blot, Expressing, Stripping Membranes, Immunofluorescence, Staining