PLCγ and IP3R-3 are involved in CVB-induced depletion of Cai2+ stores.
(A) HBMEC (left) or Caco-2 (right) cells treated with 2-APB or U73122 1 hour prior to infection (pre-treat) or 2 hours p.i. (post-treat) were infected with CVB (MOI = 1) for 14 hrs (HBMEC) or 7 hrs (Caco-2). Graph represents percentage of total cells expressing VP1 normalized to no-inhibitor controls (dashed line). (B) Top: Still images of HBMEC pre-treated with U73122, loaded with Fura-2 AM, and exposed to CVB (MOI = 50, t = 55 seconds). Images were pseudo colored for Cai2+ visualization. Bottom: Fluorescence intensity ratio (340/380 nm) of Fura-2AM versus time in HBMEC exposed to CVB in the absence (black) or presence of U73122 (blue). (C) Top: Still images captured at the indicated times in HBMEC transfected with control, IP3R-3, or PLCγ-1 siRNAs, loaded with Fura-2AM and exposed to CVB (MOI = 50, t = 55 seconds). Bottom: Intensity ratio plot (340/380 nm) of HBMEC loaded with Fura-2AM and transfected with control, IP3R-3, or PLCγ -1 siRNA and exposed to CVB (MOI = 50, t = 55 seconds).