Endogenous RGS14 is enriched as puncta at various cytosolic compartments of B35 cells.

<p>B35 cells were fixed and co-stained with RGS14 polyclonal antibody (<i>red</i>) and one of several organelle markers (<i>green</i>): (A) Rhodamine phalloidin, F-actin; (B) γ-tubulin, centrosomes; (C) endoplasmic reticulum, KDELR; (D) HSP60, mitochondria; (E) lysosomes; (F) Mann-6; EEA1, early endosomes; (G) GM130, Golgi apparatus; (H) α-tubulin, microtubules. Nuclei were counterstained with Hoechst (<i>blue</i>). For B35 cells co-stained with rhodamine phalloidin to label F-actin (A) and antibody against mitochondrial marker HSP60 (D), cells were fixed with 4% paraformaldehyde in 1X PBS and permeabilized in 0.1% Triton-X. B35 cells co-stained with all other organelles were fixed with 4% PFA in PHEM cytoskeleton stabilizing buffer and permeabilized with methanol. Insets represent magnified boxed regions (2x magnification), and are enlarged to the right of the merged image (detail). Scale bar, 10 μm. White arrowheads point to regions of endogenous RGS14 colocalization with mitochondrial marker HSP60 (D) and early endosome marker EEA1 (F). Cells shown are representative of approximately 600 cells observed from 40 fields of view across 3 independent experiments.