Fig 2: Treatment of H446 and H510A cells with increasing concentrations of the hGC33-PE38 immunotoxin. (A) Cytotoxicity curve for H446 cells. Dashed line represents the logarithmic dose-response logistic curve fitted to data. (B) No effect on H510A cells' survival. Dotted line represents the baseline of cytotoxicity. Cytotoxicity was calculated based on the analysis of neutral red uptake by viable cells. Mean ± SD (n=3). Statistical significance of differences vs. control (fixed value, 0) was calculated with ANOVA and Dunnett's post hoc test; *P=0.05, ***P=0.001. PE38, Pseudomonas aeruginosa exotoxin A 38; hGC33, anti-GPC3 antibody.

Fig 3: Inhibition of H446 cell proliferation treated for 48 h with increasing concentrations of the hGC33-PE38 immunotoxin. Proliferation inhibition was determined using the BrdU assay. Mean ± SD (n=3). Statistical significance of differences vs. control (point 0.0) was calculated with ANOVA and Tukey's post hoc test; *P=0.05. PE38, Pseudomonas aeruginosa exotoxin A 38; hGC33, anti-GPC3 antibody.

Fig 4: Internalisation of hGC33-PE38 immunotoxin in NCI-H446 and NCI-H510A cells visualised using confocal microscopy. Blue channel, nuclei (Hoechst 33342); green channel, live cells (calcein-AM); red channel, hGC33-PE38 (ATTO 542). (A) Intracellular localization of ATTO 542-stained hGC33-PE38 immunotoxin within NCI-H446 cells and no specific signal in NCI-H510A cells. (B) Calcein-AM staining indicates shape of the cells and tightness of cellular membrane. On the upper right corner of the right panels, non-specific binding within a dead cell is shown. Scale bar, 10 µm. Magnification, ×60. PE38, Pseudomonas aeruginosa exotoxin A 38; hGC33, anti-GPC3 antibody.

Fig 5: Graph showing a normalized estimation of the GPC3 gene expression across 67 lung cancer cell lines. Data obtained from the EMBL-EBI Expression Atlas tool. The x-axis presents expression values in FPKM. The accession number of primary dataset: E-MTAB-2706. LAD, lung adenocarcinoma; SCLC, small cell lung carcinoma; GPC3, glypican-3; FPKM, fragments per kilobase of exon model per million reads mapped.