(Bt) Cry toxins are used to control is the most important vector for the transmission of dengue fever yellow fever and other tropical diseases. toxin family (PFT) which constitute the most widespread group of toxins produced by bacteria. These toxins are soluble proteins that exert their functions by binding to specific receptors localized in the membrane of cells of susceptible organisms. After binding PFT at high toxin doses induces death by osmotic shock. However at low doses the toxin triggers defense mechanisms that allow cell survival [2]. The defense cell mechanisms brought on by small doses of PFT are less known. The endocytosis of macromolecules requires the recruitment of various proteins from the cytosol to the plasma membrane leading to invagination and subsequent excision of the membrane which forms a vacuole inside the cell. Several pathways involved in endocytosis have already been described including clathrin-mediated endocytosis (CME) caveolae phagocytosis macropinocytosis and several clathrin-independent pathways [3]. Bacteria exploit the endocytosis process to deliver PFT inside the host cells [2 4 In response infected cells have developed several mechanisms to repair the loss of integrity of the membrane caused by the PFT to counteract this strategy. This restoration capability is usually dependent on the rate and duration of the injury. Endocytosis promotes membrane sealing in response to the PFT streptolysin O and perforin in a Ca2+-dependent and dynamin-independent mechanism in kidney and HeLa cells [5]. HaCat and Cos7 cells induce endocytosis and exocytosis to survive an α-toxin in a Ca2+-impartial and dynamin-dependent mechanism [4]. A wounded membrane repair response has also been reported to seal the pore provoked by perforin. In this process endosomes and lysosomes donate membranes in a Ca2+-dependent manner [6]. Related to Bt toxins detoxification Griffitts and co-workers [7] reported that Cry5B toxin triggers an endocytic mechanism via specific receptors. This study used and rhodamine-labeled Cry5B toxin to demonstrate by fluorescence microscopy that this toxin binds to the nematode gut cells via receptors before being endocytosed [7]. Supporting that previous observation Los [8] reported that increased levels of endocytosis mediated by Rab5 and Rab11 are required to restore plasma membrane integrity in gut epithelium in TCS PIM-1 4a response to Cry5B. To date there are no reports demonstrating that Cry toxins are endocytosed in insect cells or whether the endocytic pathway has a role in detoxification. Bacteria protein toxins affect the actin cytoskeleton using different strategies. A group of toxins such as the binary and large clostridial TCS PIM-1 4a glucosylating toxin and the Tc toxins of directly target the actin molecule [9]. Another group interacts with actin-binding proteins to regulate actin cytoskeleton function during internalization [10]. Pore forming toxins can interact directly with TCS PIM-1 4a actin to enhance actin polymerization [11] or indirectly MRC1 to promote toxin oligomerization and endocytosis [12]. Interestingly it has been identified that actin can bind to Cry in Lepidopteran and Dipteran larvae [13 14 Based on proteomics studies it has been reported that Cry toxins affect actin accumulation in and [14 15 The proteomic profile study showed that actin protein family members are differentially up- or down-regulated in response to Cry11Aa intoxication. One of these actin genes (Accession Number: AAEL005961) was upregulated two times after treatment with sub-lethal doses of Cry11Aa toxin in larvae. Based on those results it has been suggested that actin may have a role in the toxin mode of action [16]. Here we characterized the endocytic TCS PIM-1 4a mechanism brought on by sub-lethal doses of Cry11Aa and Cry1Ab toxins that are active against Diptera and Lepidoptera respectively in an Mos20 cell line. Our results showed that Mos20 cells internalized both toxins independently of their specificity. This finding suggests that endocytosis is usually a general mechanism that insect cells use to cope with pore forming toxins independently of their toxicity. This general endocytic mechanism is usually mediated by clathrin and flotillin. Our results also exhibited that.