Fig. 6

Characterization of liver stage EEF from Indian P. vivax strains. a Representative images of P. vivax infected HCO4 cells stained with anti UIS4 antibody, 5 days post infection. EEF’s of different sizes are seen. b Quantification of EEF area from HCO4 cells infected with P. vivax sporozoites and fixed days 5 post infection. Datapoints represent individual EEF, results pooled from multiple wells of the same infection. a–e, represent the area distribution from three independent infections. a–d is from HCO4 cells, e is from primary human hepatocytes. Dashed line is at 78 μm², fraction of parasites lesser than 78 μm² per infection is indicated in the boxed area. c Reduction in the EEF numbers upon drug treatment. P. vivax sporozoite infected HCO4 cells are treated with indicated compounds daily for 4 days post infection, fixed, immunostained for pvUIS4 and number of EEF’s assessed. Data pooled from 30 wells of 384 well plate per condition. ‘MMV1’ denotes MMV390048, a PI4K inhibitor with known activity against liver stage P. vivax forms. Results averaged from three independent infections. Data normalized to DMSO of each infection, error bars indicate standard deviations. dP. vivax PVM is positive for LC3. P. vivax infected HCO4 cells were immunostained for UIS4 and the host autophagy marker LC3. EEF’s corresponding to both small and large forms from the same infection are shown. Results representative of at least three independent infections. Scale bar is 20 μm. e Scatter plot showing the intensity of LC3 per EEF spanning different size ranges of EEF shows a linear relationship. Corr corresponds to correlation value