Fig. 6.

Coimmunoprecipitation and confocal microscopic evidence for association of ACL with PHP in INS 832/13 cells. A: INS 832/13 cells were were incubated in the presence of either basal (2.5 mM; LG) or HG (20 mM) for 45 min. Samples were immunoprecipitated with ACL, as described in materials and methods. The immunoprecipitates (IP) were resolved by SDS-PAGE and probed with PHP antibody. A representative Western blot (WB) from 2 studies is shown. B: quantification of PHP levels from 2 studies, as described above. C: INS 832/13 cells were cultured on a coverslip for 24 h and then maintained overnight in low-glucose/low-serum medium. Cells were further incubated in the presence of either LG (2.5 mM; i and ii) or HG (20 mM; iv and v) for 45 min. Localization of PHP and ACL proteins was determined using respective antibodies (see materials and methods for additional details). PHP was stained green, whereas ACL was stained red. C, iii: merged images of i and ii; C, vi: merged images of iv and v. Data are representative of 2 experiments. D: INS 832/13 cells were cultured on a coverslip for 24 h and then maintained overnight in low-glucose/low-serum medium. Cells were incubated further in the presence of either LG (2.5 mM; i and ii) or HG (20 mM iv and v) for 45 min. Localization of nm23-H1 and ACL proteins was determined using respective antibodies (see materials and methods for additional details). nm23-H1 was stained green, whereas ACL was stained red. D, iii: merged images of i and ii; D, vi: merged images of iv and v. Data are representative of 2 experiments.