Journal: International Journal of Molecular Sciences

Article Title: Insight into the Interactome of Intramitochondrial PKA Using Biotinylation-Proximity Labeling

doi: 10.3390/ijms21218283

Figure Lengend Snippet: Protein kinase A (PKA) catalytic subunit α 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 α-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 α-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.

Article Snippet: Protein immunodetection was performed using primary antibodies directed against PKA C-α (Cell Signaling; 4782S), PKA RI-α/β (Cell Signaling; 3927), Phospho-PKA Substrate (Cell Signaling; 9624S), α-Tubulin (Cell Signaling; 2144) SDHa (Abcam; ab14715), SOD2 (Santa Cruz; sc-30080), NDUF9a (Abcam; ab14713), Cytochrome c (Abcam; ab110325), TOM20 (Santa Cruz; (F-10) sc-17764), UQCRC2 (Abcam; ab14745), PAM16 (Abcam; Ab184157), Hsp60 (Santa Cruz; sc-13115), Hsp70 (Santa Cruz; sc-24), V5 (Cell Signaling; 13202S), Myc (Cell Signaling; 2276S), ATP5a (Abcam; ab14730), MFN2 (Cell signaling; 9482S), ERp57 (R&Dsystems; AF8219).

Techniques: Western Blot, Isolation, Expressing, Plasmid Preparation, Marker

Journal: Journal of Inflammation Research

Article Title: Significance of Immunogenic Cell Death-Related Prognostic Gene Signature in Cervical Cancer Prognosis and Anti-Tumor Immunity

doi: 10.2147/JIR.S410140

Figure Lengend Snippet: Expression of the PDIA3 gene in cervical cancer (CC) and its prognostic value. ( A ) The cell types of IPGs distribution in the single-cell analysis of the GSE168652 database were described. ( B ) UMAP visualization of PDIA3 in major cell subpopulations of cervical cancer. ( C ) The expression levels of HPV16- and HPV18-positive groups were compared with normal groups in the GSE6791 database (* p < 0.05, ** p < 0.01, and *** p < 0.001). ( D ) Cox survival analysis showing hazard ratios for different IPGs. ( E ) Differences in PDIA3 expression between normal and cancer tissue samples (* p < 0.05). ( F and G ) Kaplan–Meier curves showing differences in overall survival ( F ) and progression-free survival ( G ) between PDIA3 high and low expression groups; ( H – J ) PDIA3 protein expression in normal cervical tissue samples and CC tissue samples. Normal cervical tissue ( H ), cervical squamous cancer ( I ), and cervical adenocarcinoma tissue ( J ).

Article Snippet: PDIA3 antibody (Proteintech, Wuhan, China), GAPDH antibody (CST, Shanghai, China), goat anti-rabbit IgG antibody secondary antibody (Zsbio, Beijing, China), and goat anti-mouse IgG H&L/Cy3 (Beyotime, Shanghai, China).

Techniques: Expressing, Single-cell Analysis

Journal: Journal of Inflammation Research

Article Title: Significance of Immunogenic Cell Death-Related Prognostic Gene Signature in Cervical Cancer Prognosis and Anti-Tumor Immunity

doi: 10.2147/JIR.S410140

Figure Lengend Snippet: PDIA3 is highly expressed in cervical cancer (CC) tissues and cell lines. ( A and B ) Western blot analysis reveals a high expression of PDIA3 in CC cell lines. ( C ) Distribution of PDIA3 in CC cell lines. ( D ) Immunohistochemistry reveals PDIA3 expression in CC tissues, with normal cervical tissue serving as a positive control (200×). ( E ) IPGs mRNA expression relative to GAPDH in cervical cancer samples comparing (* p < 0.05, ** p < 0.01, and *** p < 0.001).

Article Snippet: PDIA3 antibody (Proteintech, Wuhan, China), GAPDH antibody (CST, Shanghai, China), goat anti-rabbit IgG antibody secondary antibody (Zsbio, Beijing, China), and goat anti-mouse IgG H&L/Cy3 (Beyotime, Shanghai, China).

Techniques: Western Blot, Expressing, Immunohistochemistry, Positive Control

Journal: eLife

Article Title: Co-movement of astral microtubules, organelles and F-actin by dynein and actomyosin forces in frog egg cytoplasm

doi: 10.7554/eLife.60047

Figure Lengend Snippet:

Article Snippet: The ER was also probed with labeled anti-Protein disulfide-isomerase A3 (PDIA3) (Boster Bio #PB9772).

Techniques: Labeling, Recombinant, Expressing, Software