Immunocytochemical staining of HeLa cells using Caldesmon 1 antibody (R020823, 1:1,000), Top panel: wild-type (WT); Bottom panal: Caldesmon 1 shRNA knockdown (KD). Nuclei were stained blue with DAPI; Caldesmon 1 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.

Validation of Caldesmon 1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Caldesmon 1 antibody (R020823, 1:2,000) and analyzed using BD flow cytometer.

Flow cytometric analysis of caldesmon 1 expression in H9c2 cells using caldesmon 1 antibody (R020823, 1:2,000). Green, isotype control; red, caldesmon 1.

Immunocytochemical staining of H9C2 cells with Caldesmon 1 antibody (R020823, 1:1,000) . Nuclei were stained blue with DAPI; Caldesmon 1 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain：Medium. Scale bar, 20 μm.

Western blotting analysis using caldesmon 1 antibody (R020823). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with caldesmon 1 antibody (R020823, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using caldesmon 1 antibody (R020823). Caldesmon 1 expression in wild type (WT) and caldesmon 1 (CALD1) shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with caldesmon 1 antibody (R020823, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.