Figure 2.

Inhibition of bystander effect mediated by C01 and C02. (A-a) Time course of rPMD2-induced calcium channel formation. Erythrocytes were loaded with Fluo-4 AM. Sub lytic concentration of rPMD2 (50 ng) was added and the increase in calcium was monitored by confocal microscopy. Selected pairs of DIC and fluorescence images with time elapsed between frames in seconds (Sec) are shown. In the case of PMI treatment, C01 or C02 were added along with rPMD2. (A-b) (i) The structures depict scaffold of (Z)-5-((1-methyl-1H-indol-3-yl)methylene)-2-thioxoimidazolidin-4-one (C01) and (ii) (Z)-5-(benzo[b]thiophen-3-ylmethylene)-2-thioxoimidazolidin-4-one (C02). (B) Phosphatidylserine exposure on the erythrocyte surface. The erythrocytes were treated with rPMD1 or rPMD2 in the presence or absence of PMIs and stained with Annexin V-FITC after 48 h. The stained erythrocytes were visualized under a confocal microscope. (C) The annexin positive erythrocytes were quantitated using a flow cytometer. (D) Average Raman spectra of the untreated erythrocytes or erythrocytes treated with rPMD2 in the presence or absence of C01 was captured using 532 nm excitation. All the Raman spectra were presented after pre-processing (baseline correction, smoothening and background removal) using asymmetric least squares smoothing method (n = 2). (E) Raman images of erythrocytes were observed at 1,131 cm⁻¹ which demonstrates the distribution of methemoglobin (n = 2).