Inactivation of CHIKV by INA: Inactivation of CHIKV by INA was evaluated as CPE, virus titer and viral RNA transcripts. A: BHK cells were infected with various test and control samples and virus induced CPE was observed. Cells were stained with 0.1% crystal violet dye in 4% neutral buffered formalin. No CPE was observed in cells infected with INA treated CHIKV in combination with UV irradiation and cells replicated normally as evident by the stain uptake by the cells similar to control cells without virus infection. Irradiation of CHIKV alone and treatment with DMSO (since INA stocks were made in DMSO) did not affect the infectivity as evident by the loss of cells in these wells. B: Virus titer was measured in the supernatant of the cells infected with non-inactivated and INA-inactivated CHIKV. No virus was detected in the supernatant of INA-inactivated CHIKV and treatment with INA alone did not affect the virus replication. C: Viral transcripts were detected in the supernatants of cells infected with the non-inactivated and INA-inactivated CHIKV. No viral transcripts were detected in the cells infected with CHIKV inactivated with 50, 100 and 200 μM dose of INA in combination with UV irradiation. D: Western Blot analysis of the CHIKV proteins showed low binding of anti-CHIKV polyclonal antibody to CHIKV upon inactivation with INA. 1 = CHIKV; 2 = CHIKV + UV Irradiation; 3 = CHIKV + INA (200 μM); 4 = CHIKV + INA (50 μM) + UV Irradiation; 5 = CHIKV + INA (100 μM) + UV Irradiation; 6 = CHIKV + INA(200 μM) + UV Irradiation; and 7 = INA (200 μM) alone. Irr = UV irradiation.