Fig. 4
From: Novel γ-sarcoglycan interactors in murine muscle membranes
NKCC1 co-localizes with the sarcoglycan complex in large, detergent-stable vesicular structures. A, B HA-CFP-tagged NKCC1 (panels a, a’, blue in merges) co-localizes in punctae visualized with both A our affinity-purified anti-Sgcg and B anti-Sgcg from Proteintech Group (PTG) (panels b, b’, red in merges) after expression in RH30 rhabdomyosarcoma cells with Flag-tagged Sgcg, Sgcd, and Sgcb. Enlargements shown (a’–d’) of the boxed areas in panels a–d. C Specificity control with nonimmune rabbit (Rb) IgG substituted for anti-Sgcg. D Signal overlaps of HA-CFP-NKCC1 (panels a, a’, blue in merges) and the sarcoglycan complex (panels a, a’, red in merges) persist even when cells are briefly extracted with 0.1% Triton X-100 before fixation. E The HA-CFP tag alone (panel a, blue in merge) does not persist in pre-extracted RH30 cells co-transfected with Flag-tagged Sgcg, Sgcd, and Sgcb (panel a, red in merge). A–E Phalloidin-stained actin filaments delineate cell boundaries (panels c, c’, green in merges). Bars (d), 20 μm; bars (d’), 5 μm. Overlapping HA and Flag signals in magenta (arrows)