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Fig. 3. Pontin and Reptin form a complex with Hint1 in HEK293 cells. (A) HEK293 cells were transiently transfected with FLAG-tagged Pontin, FLAG-tagged Reptin and myc₆-tagged Hint1 alone or in combination. Protein complexes were immunoprecipitated with anti-FLAG M2 monoclonal antibodies and analysed on western blots with anti-myc (9E10) monoclonal antibody. (B) In opposite type of experiments where cells were transfected with FLAG-tagged Hint1, myc₆-tagged Pontin or myc₆-tagged Reptin, co-precipitation was only detectable when cells were cotransfected with both Pontin-FLAG or Reptin-FLAG and Hint1-myc₆ (lanes 8 and 10). After cotransfection of Hint1-FLAG and Hint1-myc₆, Hint1 dimers were detected in western blots (lane 11). (C) Association of endogenous Hint1 and Pontin in HEK293 cell lysates as shown in co-immunoprecipitation experiments with anti-Pontin ( ${alpha}$ -23) antibody and western blot detection with anti-Hint1 ( ${alpha}$ -21). **Heavy and *light chain of the precipitating antibody.

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