Human NSCLC platelets express and release elevated TLT-1 and display a distinct phenotype. (A) Representative FC histograms of isolated platelets from NSCLC patients and controls for surface TLT-1 (n = 24 for NSCLC, n = 15 for control), with their respective quantitation of median fluorescence intensities (Median FI) in the right panel (statistical significance tested by Mann–Whitney test). (B) Platelet TLT-1 mRNA levels in controls and NSCLC patients shown as the respective RT-PCR bands on agarose gel (upper panel); the band size is indicated by bp. (C) The immunoblot images show comparison of TLT-1 total protein expression in platelets from controls or NSCLC patients on Western blot, with β-tubulin (β-tub) as loading control as shown. (D) Representative FC histograms (left) of isolated platelets from NSCLC patients and controls for surface activation receptor (αIIb-β3), and respective quantitation (right) measured by PAC1 assay (n = 24 for NSCLC, n = 15 for control). Statistical significance tested by Mann–Whitney test. (E) P-sel surface expression with representative FC pseudocolor plots (left) and quantitation (right) of washed platelets from NSCLC patients and controls (n = 15 for control and n = 24 for NSCLC). Statistical significance tested by Mann–Whitney test. (F) Representative transmission EM micrographs for control and NSCLC platelets (n = 4 in each group) shown (scale bar, 1 µm) from samples from each group. The washed platelets were fixed and negatively stained as described in methods. (G) Quantitation soluble TLT-1 in plasma in control (blue) and NSCLC groups (statistical significance tested by Mann–Whitney test). (H) Activated and adhered platelets from NSCLC or control groups as observed by confocal microscopy showing TLT-1 being released by NSCLC platelets. Representative micrographs are shown with zoomed in portion of merged image on the extreme right. The scale bar represents 15 µm. The data are presented as scatter plots indicating mean ± SEM. The P values shown correspond to non-parametric Mann–Whitney test. All datasets represent a minimum of three independent experiments. Significant differences are indicated with asterisks (*, P < 0.05; ***, P < 0.001). Source data are available for this figure: SourceData F1.
Figure 1.

Human NSCLC platelets express and release elevated TLT-1 and display a distinct phenotype. (A) Representative FC histograms of isolated platelets from NSCLC patients and controls for surface TLT-1 (n = 24 for NSCLC, n = 15 for control), with their respective quantitation of median fluorescence intensities (Median FI) in the right panel (statistical significance tested by Mann–Whitney test). (B) Platelet TLT-1 mRNA levels in controls and NSCLC patients shown as the respective RT-PCR bands on agarose gel (upper panel); the band size is indicated by bp. (C) The immunoblot images show comparison of TLT-1 total protein expression in platelets from controls or NSCLC patients on Western blot, with β-tubulin (β-tub) as loading control as shown. (D) Representative FC histograms (left) of isolated platelets from NSCLC patients and controls for surface activation receptor (αIIb-β3), and respective quantitation (right) measured by PAC1 assay (n = 24 for NSCLC, n = 15 for control). Statistical significance tested by Mann–Whitney test. (E) P-sel surface expression with representative FC pseudocolor plots (left) and quantitation (right) of washed platelets from NSCLC patients and controls (n = 15 for control and n = 24 for NSCLC). Statistical significance tested by Mann–Whitney test. (F) Representative transmission EM micrographs for control and NSCLC platelets (n = 4 in each group) shown (scale bar, 1 µm) from samples from each group. The washed platelets were fixed and negatively stained as described in methods. (G) Quantitation soluble TLT-1 in plasma in control (blue) and NSCLC groups (statistical significance tested by Mann–Whitney test). (H) Activated and adhered platelets from NSCLC or control groups as observed by confocal microscopy showing TLT-1 being released by NSCLC platelets. Representative micrographs are shown with zoomed in portion of merged image on the extreme right. The scale bar represents 15 µm. The data are presented as scatter plots indicating mean ± SEM. The P values shown correspond to non-parametric Mann–Whitney test. All datasets represent a minimum of three independent experiments. Significant differences are indicated with asterisks (*, P < 0.05; ***, P < 0.001). Source data are available for this figure: SourceData F1.

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