ll 24  (ATCC)

Journal: Scientific Reports

Article Title: ACPA prevents lung fibroblast-to-CAF transformation by reprogramming the tumor microenvironment through NSCLC-derived exosomes

doi: 10.1038/s41598-025-29726-4

Figure Lengend Snippet: Control and IC50 ACPA-applied NSCLC cell exosomes were successfully characterized. a , b Micrographs representing phase-contrast (PC, scale bar, 50 μm) view of ( a ) A549 NSCLC and ( b ) LL24 healthy human lung fibroblasts, 400x. ( c ) Flow cytometric analysis of tetraspanin CD9, CD63 and CD81 markers in control and ACPA-applied A549 cell exosomes. Fluorescence (CD9-APC, CD63-FITC and CD81-PE) on the x-axis vs. number of events (Count (%)) on the y-axis ( n = 3). d–f TEM analysis. ( d ) TEM images and ( e , f ) size distribution diameters from TEM images of control and ACPA-applied A549 cell exosomes ( n = 20). g–j Representative NTA of ( g , h ) control and ( i , j ) ACPA-applied A549 cell exosomes under 100x dilutions. Graphs g and i depict the finite track length adjustment (FTLA) size per concentration as triplicates and graphs h and j depict the average FTLA size per concentration.

Article Snippet: A549 non-small cell lung adenocarcinoma cell line A549 (CCL- 185) and LL24 healthy lung fibroblasts (CCL-151) (all from ATCC, USA) were cultured with DMEM High Glucose and Ham’s F12K, respectively, supplemented with 10–15% fetal bovine serum (FBS, Capricorn, USA), or 2% L-glutamine (Capricorn, USA) and 1% penicillin-streptomycin (Capricorn, USA).

Techniques: Control, Fluorescence, Concentration Assay

Journal: Scientific Reports

Article Title: ACPA prevents lung fibroblast-to-CAF transformation by reprogramming the tumor microenvironment through NSCLC-derived exosomes

doi: 10.1038/s41598-025-29726-4

Figure Lengend Snippet: ACPA-applied NSCLC cell exosomes at a lower dose diminish LL24 HLFs compared to control within 12 h. ( a ) qRT-PCR analysis for relative miRNA fold change values of miR-21, miR-23a and miR-23b relative to U6 gene ( n = 4). b–d Proliferation indices of LL24 HLFs following the application of control or ACPA-applied A549 NSCLC cell exosomes at doses of 10, 50 and 100 µg/ml for ( b ) 1, ( c ) 2 or ( d ) 3 days by MTT assay in mean ± SEM ( n = 5, * p < 0.05 by one-way analysis of variance (ANOVA) and post-hoc Duncan’s test). All proliferation data are presented in absorbance (A570 nm). e–h Real-time normalized proliferation indices of ( e ) control or ACPA-applied A549 NSCLC cell exosome-administered HLFs presented with control and exosomes at doses of 10, 50 and 100 µg/ml along with the IC50 graphs of ( f ) control and ( g ) ACPA-applied A549 NSCLC cell exosomes for 4 days ( n = 3 for exosome-applied groups and n = 6 for controls) and ( h ) LL24 fibroblasts following co-culture with A549 cells at ratio of 1:1, 1:5 or 5:1 ( n = 6). ( i ) Treatment response time points of LL24 fibroblasts following co-culture with A549 cells when cell index (CI) is at 50 or 80%.

Article Snippet: A549 non-small cell lung adenocarcinoma cell line A549 (CCL- 185) and LL24 healthy lung fibroblasts (CCL-151) (all from ATCC, USA) were cultured with DMEM High Glucose and Ham’s F12K, respectively, supplemented with 10–15% fetal bovine serum (FBS, Capricorn, USA), or 2% L-glutamine (Capricorn, USA) and 1% penicillin-streptomycin (Capricorn, USA).

Techniques: Control, Quantitative RT-PCR, MTT Assay, Co-Culture Assay

Journal: Scientific Reports

Article Title: ACPA prevents lung fibroblast-to-CAF transformation by reprogramming the tumor microenvironment through NSCLC-derived exosomes

doi: 10.1038/s41598-025-29726-4

Figure Lengend Snippet: ACPA-applied NSCLC cell exosomes at a lower dose inhibited PDPN, FAP and α-SMA expressions in HLFs compared to control. Representative univariate histograms ( a–c ) and bar graphs ( d–f ) of FCM depicting the expressions of ( a , d ) PDPN, ( b , e ) FAP and ( c , f ) α-SMA in LL24 fibroblasts following control or ACPA-applied A549 NSCLC cell exosomes at doses of 100–50 µg/ml, respectively ( n = 3). g-h Secretome profile of LL24 fibroblasts including ( g ) IL-6 and ( h ) IL-8 on days 1, 2 and 3 following the application of control or ACPA-applied A549 NSCLC cell exosomes at doses of 10, 50 and 100 µg/ml by ELISA. ( n = 3, * p < 0.05 by one-way analysis of variance (ANOVA) and post-hoc Duncan’s test).

Article Snippet: A549 non-small cell lung adenocarcinoma cell line A549 (CCL- 185) and LL24 healthy lung fibroblasts (CCL-151) (all from ATCC, USA) were cultured with DMEM High Glucose and Ham’s F12K, respectively, supplemented with 10–15% fetal bovine serum (FBS, Capricorn, USA), or 2% L-glutamine (Capricorn, USA) and 1% penicillin-streptomycin (Capricorn, USA).

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: ACPA prevents lung fibroblast-to-CAF transformation by reprogramming the tumor microenvironment through NSCLC-derived exosomes

doi: 10.1038/s41598-025-29726-4

Figure Lengend Snippet: ACPA-applied NSCLC cell exosomes reduce fibroblast growth via carbohydrate, lipid and amino acid metabolic pathways. ( a–b ) PCA plots, ( c ) VIP scores, ( d ) heat maps and ( e–i ) mean-standard deviation graphs of metabolites of LL24 fibroblasts following treatment of ACPA-applied and control NSCLC cell exosomes and control medium containing exosome-depleted FBS ( n = 3, * p < 0.05 by one-way analysis of variance (ANOVA) and post-hoc Duncan’s test).

Article Snippet: A549 non-small cell lung adenocarcinoma cell line A549 (CCL- 185) and LL24 healthy lung fibroblasts (CCL-151) (all from ATCC, USA) were cultured with DMEM High Glucose and Ham’s F12K, respectively, supplemented with 10–15% fetal bovine serum (FBS, Capricorn, USA), or 2% L-glutamine (Capricorn, USA) and 1% penicillin-streptomycin (Capricorn, USA).

Techniques: Standard Deviation, Control