Western blot for Igl shRNA transfectants. A representative Western blot is shown with one biological replicate each for the GFP control shRNA transfectant, the Igl1-specific (272–300), the Igl (1198–1226), the Igl (2412–2440), and the Igl (2777–2805) shRNA transfectants. HM1:IMSS samples are not shown. Results shown are representative of three biological replicates per shRNA transfectant with each sample run in triplicate. Serial dilutions of the crude lysates (1:2, 1:4, and 1:8) were also performed for each sample. Each membrane was probed with anti-actin antibody as a loading control, or with anti-Igl1 antibody. Igl1 protein levels for the Igl shRNA and GFP shRNA transfectants and HM1:IMSS nontransfected amebae are summarized in Table 4.