Human ERp57/PDIA3 Antibody

Detection of ERp57/PDIA3 by Western Blot

Protein kinase A (PKA) catalytic subunit alpha is localized within mitochondria of HeLa cells. (A) Representative immunoblots (n = 3) of PKA catalytic (cat) subunit, PKA regulatory (reg) subunit, the mitochondrial protein SDHa and of the cytosolic protein alpha -Tubulin with corresponding total protein load (TPL) in total cell lysate (TCL), cytosolic (Cyto) and mitochondrial (Mito) fractions isolated from HeLa cells. (B) Representative immunoblots (n = 3) of PKA cat, PKA reg, SDHa and TOM20 with corresponding total protein load (TPL) in mitochondrial fractions isolated from HeLa cells and treated as indicated. (C) Representative micrographs (n = 6) of HeLa cells expressing empty vector (pcDNA), PKA-Myc or mt-PKA-Myc and labelled with anti-Myc (green) and anti-TOM20 (red) as a mitochondrial marker. Scale bars = 5 μm. (D) Representative immunoblots (n = 3) of Myc, SDHa and alpha -Tubulin with corresponding total protein load (TPL) in TCL, cyto and mito fractions isolated from HeLa cells expressing empty vector (pcDNA), PKA-Myc or mt-PKA-Myc. (E) Representative immunoblots (n = 3) of Myc, TOM20, SDHa and SOD2 with corresponding total protein load (TPL) in mitochondrial enriched fractions isolated from HeLa cells expressing empty vector (pcDNA), PKA-Myc or mt-PKA-Myc and treated as indicated. (F) Representative immunoblots (n = 3) of Myc, PKA Cat, ATP5a, MFN2 and ERp57 with corresponding TPL in pellet and supernatant (SN) obtained after treatment of mitochondria isolated from HeLa cells expressing PKA-Myc or mt-PKA-Myc and treated as indicated. (G) Representative immunoblots (n = 5) of phospho-PKA substrates and Myc with corresponding total protein load (TPL) in mitochondrial enriched fractions isolated from HeLa cells expressing empty vector (pcDNA), PKA-Myc or mt-PKA-Myc. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33167377), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of ERp57/PDIA3 by Western Blot

Mitochondrial matrix-localized Src regulates mitochondrial shape. a Representative immunoblotting (n = 3) of Src, cytosolic tubulin and mitochondrial SDHA in total cell lysate (TCL), cytosol- (cyto) and mitochondria- (mito) enriched fractions of Src+/+ and Src−/− MEFs, HEK293 and HeLa. b Representative immunoblotting (n = 3) of Src, the IMM protein SDHA and the OMM protein TOM20 in mitochondria-enriched fractions of HeLa cells treated as indicated. c Representative immunoblotting (n = 4) of Src, Smac-diablo, ATP5a and ERp57 in supernatant (SN) and pellet obtained from mitochondria-enriched fractions of HeLa cells treated with digitonin as indicated. d Representative micrographs (n = 3) of the plasma membrane marker wheat germ agglutinin (WGA), V5 and TOM20 labeling in Src+/+ and Src−/− mouse embryonic fibroblasts (MEFs) expressing pcDNA, Src-V5, mitochondria-targeted mtSrc-V5 and plasma membrane-targeted pmSrc-V5. e Quantitative analysis of mitochondrial morphology in region of interest (ROI) as shown in c. Scale bars: 20 μm. Data are presented as mean ± S.E.M. Data with different letters are statistically different (p < 0.05), according to one-way ANOVA followed by Tukey’s post hoc test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35637383), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of ERp57/PDIA3 by Western Blot

Mitochondrial matrix-localized Src regulates mitochondrial shape. a Representative immunoblotting (n = 3) of Src, cytosolic tubulin and mitochondrial SDHA in total cell lysate (TCL), cytosol- (cyto) and mitochondria- (mito) enriched fractions of Src+/+ and Src−/− MEFs, HEK293 and HeLa. b Representative immunoblotting (n = 3) of Src, the IMM protein SDHA and the OMM protein TOM20 in mitochondria-enriched fractions of HeLa cells treated as indicated. c Representative immunoblotting (n = 4) of Src, Smac-diablo, ATP5a and ERp57 in supernatant (SN) and pellet obtained from mitochondria-enriched fractions of HeLa cells treated with digitonin as indicated. d Representative micrographs (n = 3) of the plasma membrane marker wheat germ agglutinin (WGA), V5 and TOM20 labeling in Src+/+ and Src−/− mouse embryonic fibroblasts (MEFs) expressing pcDNA, Src-V5, mitochondria-targeted mtSrc-V5 and plasma membrane-targeted pmSrc-V5. e Quantitative analysis of mitochondrial morphology in region of interest (ROI) as shown in c. Scale bars: 20 μm. Data are presented as mean ± S.E.M. Data with different letters are statistically different (p < 0.05), according to one-way ANOVA followed by Tukey’s post hoc test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35637383), licensed under a CC-BY license. Not internally tested by R&D Systems.