anti pdia3 rabbit polyclonal antibody  (Proteintech)

Journal: International Journal of Molecular Sciences

Article Title: Robust CXCL10/IP-10 and CCL5/RANTES Production Induced by Tick-Borne Encephalitis Virus in Human Brain Pericytes Despite Weak Infection

doi: 10.3390/ijms25147892

Figure Lengend Snippet: TBEV-infected HBVP. ( A ) Virus titers in cell-free supernatants of infected HBVP were evaluated on days 0, 1, 3, 6 and 10 p.i. by plaque assay. The results represent the mean ± SEM of 3 independent experiments that were run in biological triplicates. ( B ) The number of intracellular TBEV RNA copies was evaluated on days 1, 3, 6 and 10 p.i. by qRT-PCR. The results represent the mean ± SEM of 2 independent experiments that were run in biological triplicates. ( C ) HBVP were infected with TBEV strain Hypr and examined for signs of CPE using light microscopy (scale: 100 µm). ( D ) HBVP grown on slides were infected with the TBEV strains Hypr and Neudoerfl. At 3 days p.i., cells were fixed and stained with anti-PDIA3 (red) and anti-flavivirus envelope protein antibody (green). Nuclei counterstained with DAPI are shown in blue (scale: 20 µm).

Article Snippet: Cells were then double-labeled with either rabbit anti-PDGFRβ monoclonal antibody (1:100, MA5-15143, Invitrogen) and mouse anti-flavivirus group antigen monoclonal antibody (1:250, MAB10216, Invitrogen) or rabbit anti-PDIA3 polyclonal antibody (1:250, HPA003230, Sigma-Aldrich, St. Louis, MO, USA) together with anti-flavivirus group antigen antibody.

Techniques: Infection, Virus, Plaque Assay, Quantitative RT-PCR, Light Microscopy, Staining

Journal: Viruses

Article Title: PDIA4 Is a Host Factor Important for Lymphocytic Choriomeningitis Virus Infection.

doi: 10.3390/v15122343

Figure Lengend Snippet: Figure 2. The PDIA4 expression level is upregulated by LCMV infection both in vitro and in vivo. (A) The expression level of LCMV-NP in A549 cells at 12, 24, 36, and 48 h.p.i. determined by RT-PCR. (B) The expression level of detectable PDIs in A549 cells at 12, 24, 36, and 48 h.p.i. determined by RT-PCR. (C) The protein expression level of PDIA3, PDIA4, PDIA5, and PDIA6 in mock-/LCMV- infected A549 cells at 12, 24, 36, and 48 h.p.i. determined by Western blot. (D) The protein expression level of PDIA3, PDIA4, PDIA5, and PDIA6 in mock-/LCMV-infected HEK293T cells at 12, 24, 36, and 48 h.p.i. determined by Western blot. (E) The protein expression level of PDIA3, PDIA4, PDIA5, and PDIA6 in mock-/LCMV-infected NHBE cells at 48 h.p.i. determined by Western blot. (F) The relative mRNA expression level of PDIA4 in the kidney and lung of mock-/LCMV-infected mice (relative to mock, n = 5, unpaired t test). *** p < 0.001, **** p < 0.0001.

Article Snippet: The anti-PDIA3 rabbit polyclonal antibody (15967-1-AP), antiPDIA4 rabbit polyclonal antibody (14712-1-AP), anti-PDIA5 rabbit polyclonal antibody (15545-1-AP), anti-V5 antibody (14440-1-AP), horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG(H+L) (SA00001-2), and HRP-conjugated goat anti-mouse IgG(H+L) (SA00001-1) were purchased from Proteintech Group (Wuhan, China).

Techniques: Expressing, Infection, In Vitro, In Vivo, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: PDIA3 expression is up-regulated in OSCC tissues. (A and B) The mRNA expression of PDIA3 in the normal and OSCC tissues in the OSCC dataset from TCGA. (A) Compare all tumor and normal tissues. (B) Only compare paired tumors and normal tissues. (C) The protein expression of PDIA3 by IHC was retrieved from the HPA database. (D) The immunofluorescent staining of PDIA3 in human OSCC tissues and non-tumorous tissues. Left, representative images; right, quantification of the intensity of the PDIA3 staining. Bar = 20 μm. (E) The IHC staining of PDIA3 in human OSCC tissues and normal tissues. Left, representative IHC images. Upper: bar = 200 μm; lower: bar = 50 μm. Right, quantification of the intensity of the PDIA3 staining. (F) PDIA3 mRNA expression in normal tissues and tumors at different T stages. (G) PDIA3 level in normal tissues and OSCC tumors at different N stages. ***, P < 0.001. A P < 0.05 is significantly different.

Article Snippet: The slides were then incubated overnight at 4 °C with primary rabbit polyclonal anti-PDIA3 antibody (1:300; Proteintech, USA).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: The diagnostic and prognostic value of PDIA3 in OSCC. The PDIA3 expression and clinical data were retrieved from the OSCC dataset in TCGA. (A) The ROC curve for analyzing the value of PDIA3 in the diagnosis of OSCC. (B–D) The Kaplan-Meier method was used to analyze the value of PDIA3 in predicting the prognosis of OSCC patients. The Kaplan-Meier curves for (B) OS, (C) DSS and (D) PFI are shown here. The Logrank test was used to compare the curves. A P < 0.05 is significantly different.

Article Snippet: The slides were then incubated overnight at 4 °C with primary rabbit polyclonal anti-PDIA3 antibody (1:300; Proteintech, USA).

Techniques: Diagnostic Assay, Expressing, Biomarker Discovery

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: Univariate and multivariate Cox regression analysis of PDIA3 in OSCC.

Article Snippet: The slides were then incubated overnight at 4 °C with primary rabbit polyclonal anti-PDIA3 antibody (1:300; Proteintech, USA).

Techniques:

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: Down-regulation of PDIA3 inhibits the cancerous phenotypes of OSCC cells. The CAL27 and SCC25 cells were transfected with siRNA oligos targeting PDIA3 (designated as siPDIA3). Cells transfected with non-targeting oligos were used as controls (siNC). The untreated cells were also used and designated as ctrl. (A and B) Immunoblotting was performed to measure the protein level of PDIA3 in (A) CAL27 and (B) SCC25 cells. (C and D) The cell viability was measured by CCK8 assay on 24, 48, 72, and 96 h in (C) CAL27 and (D) SCC25 cells transfected with siPDIA3 or non-targeting siRNA. (E and F) The apoptosis of (E) CAL27 and (F) SCC25 cells was measured using an ELISA assay detecting the level of cytosolic nucleosomes. (G and H) The migratory ability of siRNA-transfected (G) CAL27 and (H) SCC25 cells was evaluated using a wound healing assay. Left: representative images; right: quantification of the wound healing rates. Scale bar = 100 μm *, P < 0.05, **, P < 0.01, ***, P < 0.001. A P < 0.05 is significantly different.

Article Snippet: The slides were then incubated overnight at 4 °C with primary rabbit polyclonal anti-PDIA3 antibody (1:300; Proteintech, USA).

Techniques: Transfection, Western Blot, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Wound Healing Assay

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: PDIA3 regulates the PI3K/AKT signaling pathway in OSCC. (A) The mRNA expression of PI3KCD, AKT1-3, and mTOR in OSCC and normal tissues. (B) A heatmap showing the expression levels of PI3KCD, AKT1-3, and mTOR across PDIA3 high and PDIA3 low OSCC tissues. Red: high expression level; blue: low expression level. (C) The correlation between PDIA3 and PI3KCD, AKT1-3, or mTOR was analyzed by Spearman's correlation analysis. (D and E) PDIA3 was silenced by siRNA transfection (siPDIA3). Cells transfected with non-targeting siRNA (siNC) and untreated cells (ctrl) were used as controls. The levels of phosphor-AKT and total AKT were measured by immunoblotting in (D) CAL27 and (E) SCC25 cells. *, P < 0.05, **, P < 0.01, ***, P < 0.001. A P < 0.05 is significantly different.

Article Snippet: The slides were then incubated overnight at 4 °C with primary rabbit polyclonal anti-PDIA3 antibody (1:300; Proteintech, USA).

Techniques: Expressing, Transfection, Western Blot

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: PDIA3 single-gene GSEA functional enrichment analysis. The GSEA for the differential genes related to PDIA3. (A) Enriched functions of the down-regulated genes. (B) Enriched functions of the up-regulated genes. (C) A lollipop plot showing the correlation between PDIA3 expression and immune cell infiltration in OSCC tissues by Spearman analysis. *, P < 0.05, **, P < 0.01, ***, P < 0.001, n.s, not significant. A P < 0.05 is significantly different.

Article Snippet: The slides were then incubated overnight at 4 °C with primary rabbit polyclonal anti-PDIA3 antibody (1:300; Proteintech, USA).

Techniques: Functional Assay, Expressing

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: PDIA3 expression is up-regulated in OSCC tissues. (A and B) The mRNA expression of PDIA3 in the normal and OSCC tissues in the OSCC dataset from TCGA. (A) Compare all tumor and normal tissues. (B) Only compare paired tumors and normal tissues. (C) The protein expression of PDIA3 by IHC was retrieved from the HPA database. (D) The immunofluorescent staining of PDIA3 in human OSCC tissues and non-tumorous tissues. Left, representative images; right, quantification of the intensity of the PDIA3 staining. Bar = 20 μm. (E) The IHC staining of PDIA3 in human OSCC tissues and normal tissues. Left, representative IHC images. Upper: bar = 200 μm; lower: bar = 50 μm. Right, quantification of the intensity of the PDIA3 staining. (F) PDIA3 mRNA expression in normal tissues and tumors at different T stages. (G) PDIA3 level in normal tissues and OSCC tumors at different N stages. ***, P < 0.001. A P < 0.05 is significantly different.

Article Snippet: After overnight incubation at 4 °C with rabbit polyclonal anti-PDIA3 primary antibody (1:300, Proteintech, USA), the slides were incubated with HRP-goat anti-rabbit IgG secondary antibody (Boster Biological Technology Co. Ltd, China) at 37 °C for 30 min and washed with PBS three times.

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: The diagnostic and prognostic value of PDIA3 in OSCC. The PDIA3 expression and clinical data were retrieved from the OSCC dataset in TCGA. (A) The ROC curve for analyzing the value of PDIA3 in the diagnosis of OSCC. (B–D) The Kaplan-Meier method was used to analyze the value of PDIA3 in predicting the prognosis of OSCC patients. The Kaplan-Meier curves for (B) OS, (C) DSS and (D) PFI are shown here. The Logrank test was used to compare the curves. A P < 0.05 is significantly different.

Article Snippet: After overnight incubation at 4 °C with rabbit polyclonal anti-PDIA3 primary antibody (1:300, Proteintech, USA), the slides were incubated with HRP-goat anti-rabbit IgG secondary antibody (Boster Biological Technology Co. Ltd, China) at 37 °C for 30 min and washed with PBS three times.

Techniques: Diagnostic Assay, Expressing, Biomarker Discovery

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: Univariate and multivariate Cox regression analysis of PDIA3 in OSCC.

Article Snippet: After overnight incubation at 4 °C with rabbit polyclonal anti-PDIA3 primary antibody (1:300, Proteintech, USA), the slides were incubated with HRP-goat anti-rabbit IgG secondary antibody (Boster Biological Technology Co. Ltd, China) at 37 °C for 30 min and washed with PBS three times.

Techniques:

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: Down-regulation of PDIA3 inhibits the cancerous phenotypes of OSCC cells. The CAL27 and SCC25 cells were transfected with siRNA oligos targeting PDIA3 (designated as siPDIA3). Cells transfected with non-targeting oligos were used as controls (siNC). The untreated cells were also used and designated as ctrl. (A and B) Immunoblotting was performed to measure the protein level of PDIA3 in (A) CAL27 and (B) SCC25 cells. (C and D) The cell viability was measured by CCK8 assay on 24, 48, 72, and 96 h in (C) CAL27 and (D) SCC25 cells transfected with siPDIA3 or non-targeting siRNA. (E and F) The apoptosis of (E) CAL27 and (F) SCC25 cells was measured using an ELISA assay detecting the level of cytosolic nucleosomes. (G and H) The migratory ability of siRNA-transfected (G) CAL27 and (H) SCC25 cells was evaluated using a wound healing assay. Left: representative images; right: quantification of the wound healing rates. Scale bar = 100 μm *, P < 0.05, **, P < 0.01, ***, P < 0.001. A P < 0.05 is significantly different.

Article Snippet: After overnight incubation at 4 °C with rabbit polyclonal anti-PDIA3 primary antibody (1:300, Proteintech, USA), the slides were incubated with HRP-goat anti-rabbit IgG secondary antibody (Boster Biological Technology Co. Ltd, China) at 37 °C for 30 min and washed with PBS three times.

Techniques: Transfection, Western Blot, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Wound Healing Assay

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: PDIA3 regulates the PI3K/AKT signaling pathway in OSCC. (A) The mRNA expression of PI3KCD, AKT1-3, and mTOR in OSCC and normal tissues. (B) A heatmap showing the expression levels of PI3KCD, AKT1-3, and mTOR across PDIA3 high and PDIA3 low OSCC tissues. Red: high expression level; blue: low expression level. (C) The correlation between PDIA3 and PI3KCD, AKT1-3, or mTOR was analyzed by Spearman's correlation analysis. (D and E) PDIA3 was silenced by siRNA transfection (siPDIA3). Cells transfected with non-targeting siRNA (siNC) and untreated cells (ctrl) were used as controls. The levels of phosphor-AKT and total AKT were measured by immunoblotting in (D) CAL27 and (E) SCC25 cells. *, P < 0.05, **, P < 0.01, ***, P < 0.001. A P < 0.05 is significantly different.

Article Snippet: After overnight incubation at 4 °C with rabbit polyclonal anti-PDIA3 primary antibody (1:300, Proteintech, USA), the slides were incubated with HRP-goat anti-rabbit IgG secondary antibody (Boster Biological Technology Co. Ltd, China) at 37 °C for 30 min and washed with PBS three times.

Techniques: Expressing, Transfection, Western Blot

Journal: Heliyon

Article Title: The role of PDIA3 in oral squamous cell carcinoma and its value as A diagnostic and prognostic biomarker

doi: 10.1016/j.heliyon.2023.e22596

Figure Lengend Snippet: PDIA3 single-gene GSEA functional enrichment analysis. The GSEA for the differential genes related to PDIA3. (A) Enriched functions of the down-regulated genes. (B) Enriched functions of the up-regulated genes. (C) A lollipop plot showing the correlation between PDIA3 expression and immune cell infiltration in OSCC tissues by Spearman analysis. *, P < 0.05, **, P < 0.01, ***, P < 0.001, n.s, not significant. A P < 0.05 is significantly different.

Article Snippet: After overnight incubation at 4 °C with rabbit polyclonal anti-PDIA3 primary antibody (1:300, Proteintech, USA), the slides were incubated with HRP-goat anti-rabbit IgG secondary antibody (Boster Biological Technology Co. Ltd, China) at 37 °C for 30 min and washed with PBS three times.

Techniques: Functional Assay, Expressing

Journal: Nature

Article Title: The NK cell receptor NKp46 recognizes ecto-calreticulin on ER-stressed cells

doi: 10.1038/s41586-023-05912-0

Figure Lengend Snippet: Knockout and knockdown of CALR and PDIA3. a, CRT immunoblot in WT or CALR knockout HEK293T clones compared to tubulin loading control. b, Flow cytometry of externalized CRT (Ecto-CRT) or NKp46-Ig binding on untreated HEK293T CALR−/− cells reconstituted with CALR constructs (representative histograms (left), MFI of n = 3 samples (right)). c, qRT-PCR of CALR and PDI after knockdown of indicated genes in JEG-3, normalized to 18S rRNA (n = 3 samples). d, Representative flow cytometry histograms of MHC class I surface expression (W6/32 antibody) on JEG-3, knocked down for CALR, PDI3 or with nontargeting (Ctl) siRNAs (left); MFI of 3 samples (right). e, JEG-3, knocked down for CALR and/or PDIA3 or with nontargeting (Ctl) siRNAs, were infected or not with ZIKV and analyzed for ecto-CRT by flow cytometry. Representative histograms (left); MFI of 3 samples (right). f, JEG-3, knocked down for CALR and/or PDI3 or with nontargeting (Ctl) siRNAs were infected or not with ZIKV and analyzed for NKp46-Ig binding by flow cytometry; representative histograms of NKp46-Ig binding (left) and MFI of 3 samples (right). g, Peripheral blood NK killing (n = 6 donors) of knocked down JEG-3. h, Effect of anti-CRT and/or anti-PDI on NKp46-Ig binding to JEG-3 that were infected or not with ZIKV. Representative flow histograms (left) and MFI of 3 samples (right). i, Effect of anti-CRT, anti-PDI and anti-NKp46 on peripheral blood NK killing of uninfected or ZIKV-infected JEG-3. (g,i, 4 h 51Cr release assay, E:T ratio 10:1, n = 3 donors). Data are representative of three independent experiments (a). Graphs shown mean ± SEM. Statistics were performed using one-way ANOVA (b,d—i), or two-tailed non-parametric unpaired t-test (c). P: *<0.05; **<0.01; ****<0.0001.

Article Snippet: Antibodies and reagents Antibodies and reagents used were: donkey-anti-mouse Alexa Fluor 488, LIVE/DEAD Fixable Violet Dead Cell Stain, purified mouse anti-human HLA-A, B, C (clone W6/32), Alexa Fluor 647 anti-human CD54 Antibody (clone HCD54), APC anti-human CD337 (NKp30, clone P30–15), PerCP-Cy5.5 anti-human Siglec-8 (clone 7C9), PE anti-human Siglec-7 (clone 6–434), Alexa Fluor 488 anti-Hsp70 (clone W27), anti-human CD56 -PE and -Pacific Blue (clone HCD56), CD107a-PerCP-Cy5.5 (clone H4A3), purified mouse IgG1, IgG1-FITC, IgG1-PE, IgG1-APC, IgG1-PerCP-Cy5.5, IgG1-PE-Cy7, IgG1-APC-Cy7 (clone MOPC-21), purified mouse IgG2a, IgG2a-FITC, IgG2a-Alexa Fluor 700 (clone MOPC-173), anti-mouse B220-PE (clone RA3–6B2) and CD45-PE-Cy7 (clone 30-F11) (all BioLegend); MICA/B-Alexa Fluor 700 (no. 159207) (R&D Systems); anti-mouse CD3e-PerCP (clone 145–2C11), CD8-PerCP (clone 53–6.7) and IFNγ-APC (clone XMG1.2) (all BD); anti-mouse CD8-eFluor 710 (clone 53–6.7), CD62L-APC (clone MEL-14), CD44-PE (clone IM7), CD4-eFluor 450 (clone RM4–5) and NKp46-PE (clone 29A1.4) (all eBioscience); FITC-AffiniPure goat anti-human IgG (Jackson Immunoresearch); Alexa Fluor 647 anti-mouse RAE1-γ (clone CX1, BioLegend); FITC anti-mouse H-2K b (clone AF6–88.5, BioLegend); Alexa Fluor 488 anti-phospho-Syk (clone pY348), Alexa Fluor 647 anti-phospho-CD3-zeta (clone pY142) and PE anti-phosphotyrosine (all BD Biosciences); goat anti-mouse IgG Fab recombinant secondary antibody (Invitrogen); anti-NKp46 (clone 9E2), APC anti-human Siglec-6 (clone REA852) (Miltenyi Biotec); anti-mouse CD8 mAb (clone 2.43), rabbit anti-mouse NK1.1 and anti-CSF1R, control antibody (no. LTF-2) (all BioXCell); rabbit anti-human CRT (no. ab2907, Abcam); mouse monoclonal anti-CRT (clone MAB38981, R&D Systems; FMC 75, Thermo); PDIA3 rabbit polyclonal Ab (no. A1085, Abclonal); polyclonal anti-NKp46 antibody (no. LS-C806753, LSBio); anti-human Siglec-6 (clone Mab239–90, Creative Biolabs); Alexa Fluor 488 mouse IgG1 isotype control (clone MOPC-21); PE mouse IgG1 isotype control (clone MOPC-21); Alexa Fluor 647 mouse IgG1 isotype control (clone MOPC-21), PerCP-Cy5.5 mouse IgG1 isotype control (clone MOPC-21), Alexa Fluor 488 mouse IgG2a isotype control (clone MOPC-173), Alexa Fluor 647 mouse IgG2b isotype control (BioLegend, clone MPC-11) and Pacific Blue mouse IgG2b isotype control (BioLegend, clone MPC-11).

Techniques: Knock-Out, Knockdown, Western Blot, Clone Assay, Control, Flow Cytometry, Binding Assay, Construct, Quantitative RT-PCR, Expressing, Infection, Release Assay, Two Tailed Test