Additional file 5: Figure S4. of New insights into HCV replication in original cells from Aedes mosquitoes

HCV Infection protocols. (A) for human HepaRG hepatocytes (“H”) and (B) for insect cells, Ktmos1 (“K”, Ae Aegypti) and C6/36 (“C”, Ae Albopictus). The infection was performed using HCVsp, LAT isolate, genotype 3. D, day; − before infection; D0, day of infection; D4, D7, D14, D21, D28, days post-infection and medium change. P17, P18, passages 17 and 18. HepaRG®, HepaRG cells from KIT902 (Biopredic International). Over the time, HepaRG and Ktmos1 cells in monolayer became more and more differentiated. (TIFF 300 kb

Additional file 3: Figure S2. of New insights into HCV replication in original cells from Aedes mosquitoes

Fluorescence observation of adherent Ktmos1 cells. The Ktmos1 Aedes aegypti cells were grown on thin glass (0,17 mm), 2 chambers LabTek (Nunc). The cells were fixed after different periods of cultivation with 2% PFA for 20 min at 37 °C. After permeabilization by PBS containing 0,1% Triton X100 for 2 min, the nuclei were stained by Hoechst 33,258 (Sigma). Observation was performed on motorized inverted Olympus IE81 microscope using the DIC (Differential Interference Contrast) and the DAPI filter. The panel (A) shows a late metaphase stage of a dividing cell. The panel (B) shows Ktmos1 cells in monolayer. (TIFF 925 kb

Additional file 2: Figure S1. of New insights into HCV replication in original cells from Aedes mosquitoes

Global process of the Ktmos1 cell generation from eggs hatching to the final supracellular structures. (A) Macroscopic picture showing the eggs of Aedes aegypti collected from insectary. (B) Large hollow vesicles developing at the cut ends of the larvae fragments. (C) Microscopic examination of the adherent cells and molecular identification of cells and larvae extracts by PCR targeting rDNA ITS: the upper panel shows cells in monolayer; the lower panel indicates species-diagnosis PCR of cellular samples with hollow vesicles (lane 1), adherent cells (lane 2) and “Dome-like” structures (lane 3). HEK 293 cells are the negative control, ground larvae extracts of Aedes aegypti bora bora strain are the positive control. The approximate size of the amplified product is 550 pb. (D) Microscopic examination of the hollow vesicles as supracellular structures (D1 and D2). (TIFF 751 kb

Additional file 7: Figure S6. of New insights into HCV replication in original cells from Aedes mosquitoes

Absence of HCV RNA detection in HEK 293 cells. Cells were collected at days 0 (D0), 4 (D4), 21 (D21) and 28 (D28) p.i. The inoculum HCVsp (LAT isolate, genotype 3) was used as positive control. Non-infected (mock) cells (−) and HCV-infected (+) HEK 293 cells. (TIFF 1236 kb