Figure 3. Effects of MYOC Mutations on Expression of IL1A in Stably Transduced Cell Lines MYOC expression was induced in TM-1 cell lines stably transduced with wild-type or mutant MYOC lentivirus expression constructs at 1000 ng/ml doxycycline. A parallel set of cells was left uninduced for comparison. After 6 days, conditioned media and cell lysates were collected for assay. A: Lanes of an SDS-polyacrylamide gel were equally loaded with aliquots of each sample (20 ul of cell culture media or 20 ug cell lysates) and proteins were separated by electrophoresis. Western blotting was performed with anti-MYOC antibody to quantify MYOC protein. Lysate blots were re-probed with ACTB as a control for equal gel loading. The electrophoretic mobility of molecular weight size standards is indicated. B: ELISA was used to quantify IL1A protein in equal volume samples of conditioned media.