rabbit anti hspd1  (Proteintech)

Journal: Biomedical Reports

Article Title: Heat shock protein family D member 1 mediates lung cancer cell‑induced angiogenesis of endothelial cells

doi: 10.3892/br.2025.1955

Figure Lengend Snippet: Cell migration capability of EA.hy926 cells after treatment with shHSPD1-A549 and shControl-A549 secretomes. (A) Representative images of the wound area captured at 0, 8, 16 and 24 h after secretome treatment, captured at x100 magnification. (B) Percentage of wound healing. Data are presented as mean ± SD from three independent experiments. * P<0.05. sh, short hairpin; HSPD1, heat shock protein family D member 1.

Article Snippet: The membrane was blocked with 5% (w/v) skimmed milk in phosphate-buffered saline (PBS) at room temperature for 1 h. Subsequently, the membrane was incubated overnight at 4˚C with either rabbit monoclonal anti-HSPD1 (1:1,000; cat. no. 12165; Cell Signaling Technology, Inc.) or mouse monoclonal anti-GAPDH (1:5,000; cat. no. ab8245; Abcam).

Techniques: Migration

Journal: Biomedical Reports

Article Title: Heat shock protein family D member 1 mediates lung cancer cell‑induced angiogenesis of endothelial cells

doi: 10.3892/br.2025.1955

Figure Lengend Snippet: Cell invasion of EA.hy926 cells after treatment with shHSPD1-A549 and shControl-A549 secretomes. (A) Representative images of crystal violet-stained EA.hy926 cells after 24-h treatment with secretomes, captured at x100 magnification. (B) The optical density of crystal violet-stained cells at 590 nm. Data are presented as mean ± SD from three independent experiments and expressed as a fold change relative to the untreated control. *** P<0.001. sh, short hairpin; HSPD1, heat shock protein family D member 1.

Article Snippet: The membrane was blocked with 5% (w/v) skimmed milk in phosphate-buffered saline (PBS) at room temperature for 1 h. Subsequently, the membrane was incubated overnight at 4˚C with either rabbit monoclonal anti-HSPD1 (1:1,000; cat. no. 12165; Cell Signaling Technology, Inc.) or mouse monoclonal anti-GAPDH (1:5,000; cat. no. ab8245; Abcam).

Techniques: Staining, Control

Journal: Biomedical Reports

Article Title: Heat shock protein family D member 1 mediates lung cancer cell‑induced angiogenesis of endothelial cells

doi: 10.3892/br.2025.1955

Figure Lengend Snippet: Cell aggregation of EA.hy926 cells following treatment with shHSPD1-A549 and shControl-A549 secretomes. (A) Representative images of EA.hy926 cell aggregates after secretome treatment, captured at x100 magnification. (B) Size of cell aggregates. Data are presented as mean ± SD from three independent experiments. * P<0.05, ** P<0.01. sh, short hairpin; HSPD1, heat shock protein family D member 1.

Article Snippet: The membrane was blocked with 5% (w/v) skimmed milk in phosphate-buffered saline (PBS) at room temperature for 1 h. Subsequently, the membrane was incubated overnight at 4˚C with either rabbit monoclonal anti-HSPD1 (1:1,000; cat. no. 12165; Cell Signaling Technology, Inc.) or mouse monoclonal anti-GAPDH (1:5,000; cat. no. ab8245; Abcam).

Techniques:

Journal: Biomedical Reports

Article Title: Heat shock protein family D member 1 mediates lung cancer cell‑induced angiogenesis of endothelial cells

doi: 10.3892/br.2025.1955

Figure Lengend Snippet: Endothelial tube formation of EA.hy926 cells after treatment with shHSPD1-A549 and shControl-A549 secretomes. (A) Representative images of tube formation in EA.hy926 cells after secretome treatment, captured at x40 magnification with additional zoomed-in areas. (B) Length of formed tubes. (C) Area of formed tubes. Data are presented as mean ± SD from three independent experiments. * P<0.05. sh, short hairpin; HSPD1, heat shock protein family D member 1.

Article Snippet: The membrane was blocked with 5% (w/v) skimmed milk in phosphate-buffered saline (PBS) at room temperature for 1 h. Subsequently, the membrane was incubated overnight at 4˚C with either rabbit monoclonal anti-HSPD1 (1:1,000; cat. no. 12165; Cell Signaling Technology, Inc.) or mouse monoclonal anti-GAPDH (1:5,000; cat. no. ab8245; Abcam).

Techniques:

Journal: Biomedical Reports

Article Title: Heat shock protein family D member 1 mediates lung cancer cell‑induced angiogenesis of endothelial cells

doi: 10.3892/br.2025.1955

Figure Lengend Snippet: VEGF levels in the culture supernatant of EA.hy926 cells after treatment with shHSPD1-A549 and shControl-A549 secretomes. VEGF levels were measured by ELISA. The bar graph represents mean ± SD from three independent experiments. * P<0.05, ** P<0.01. VEGF, vascular endothelial growth factor; sh, short hairpin; HSPD1, heat shock protein family D member 1.

Article Snippet: The membrane was blocked with 5% (w/v) skimmed milk in phosphate-buffered saline (PBS) at room temperature for 1 h. Subsequently, the membrane was incubated overnight at 4˚C with either rabbit monoclonal anti-HSPD1 (1:1,000; cat. no. 12165; Cell Signaling Technology, Inc.) or mouse monoclonal anti-GAPDH (1:5,000; cat. no. ab8245; Abcam).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Biomedical Reports

Article Title: Heat shock protein family D member 1 mediates lung cancer cell‑induced angiogenesis of endothelial cells

doi: 10.3892/br.2025.1955

Figure Lengend Snippet: Correlation of VEGFA expression with HSPD1 expression and overall survival in patients with lung adenocarcinoma. (A) Scatter plot showing the correlation between VEGFA and HSPD1 expression in the TCGA-LUAD cohort, analyzed using the TIMER database. (B) Kaplan-Meier survival curve comparing overall survival in lung adenocarcinoma patients with low and high VEGFA expression, obtained from the KM Plotter database. VEGF, vascular endothelial growth factor; sh, short hairpin; HSPD1, heat shock protein family D member 1; TCGA-LUAD, The Cancer Genome Atlas-Lung Adenocarcinoma; TIMER, Tumor IMmune Estimation Resource.

Article Snippet: The membrane was blocked with 5% (w/v) skimmed milk in phosphate-buffered saline (PBS) at room temperature for 1 h. Subsequently, the membrane was incubated overnight at 4˚C with either rabbit monoclonal anti-HSPD1 (1:1,000; cat. no. 12165; Cell Signaling Technology, Inc.) or mouse monoclonal anti-GAPDH (1:5,000; cat. no. ab8245; Abcam).

Techniques: Expressing

Journal: iScience

Article Title: Role of human HSPE1 for OPA1 processing independent of HSPD1

doi: 10.1016/j.isci.2023.106067

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-HSPD1 (D6F1) , Cell Signaling Technology , Cat# 12165; RRID:AB_2636980.

Techniques: Recombinant, Modification, Saline, Protease Inhibitor, Plasmid Preparation, SYBR Green Assay, cDNA Synthesis, CRISPR, Cell Culture, Sequencing, esiRNA, Negative Control, Software

Journal: iScience

Article Title: Role of human HSPE1 for OPA1 processing independent of HSPD1

doi: 10.1016/j.isci.2023.106067

Figure Lengend Snippet: OPA1 cleavage induced by HSPE1 knockdown is independent of HSPD1 (A) Western blot of HEK293T cells after individual or simultaneous knockdown of HSPD1 and HSPE1 using esiRNA. The asterisks indicate the target band. (B) Quantitation of band intensity normalized relative to GAPDH. Bars indicate averages ±SD (n = 4). (C) Relative mRNA levels of HSPE1 were determined by qPCR. Bars indicate averages ±SD (n = 8). (D) A representative band intensity of OPA1 isoforms in cells with the indicated knockdown. (E) The ratio of short isoforms of OPA1 over long isoforms is quantified ± SD (n = 4). (F) OCRs were analyzed in cells with the indicated knockdown. (G) The basal OCRs are quantified. Bars indicate averages ±SD (n = 3). (H) The maximal OCRs are quantified. Bars indicate averages ±SD (n = 3). Statistical analysis was performed by one-way ANOVA with post hoc Tukey: ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The antibodies used were HSPD1 (1:1,000 dilution, 12165, Cell Signaling), DNAJA3 (1:1,000 dilution, 11088-1-AP; Proteintech), HSPA9 (1:5,000 dilution, 14887-1-AP; Proteintech), HSPA14 (1:1,000 dilution, MA5-32413, Invitrogen), OPA1 (1:1,000 dilution, 612607; BD Biosciences), GAPDH (1:1,000 dilution, MA5-15738; Invitrogen), TOM20 (1:2,000 dilution, sc-11415; Santa Cruz Biotechnology), OMA1 (1:500 dilution, sc-515788; Santa Cruz Biotechnology), MFN1/2 (1:1,000 dilution, ab57602; Abcam), PDH (1:1,000 dilution, ab110338; Abcam), TIM23 (1:5,000 dilution, 611223; BD), and GFP (1:5,000 dilution, a gift from Dr. Peter Devreotes).

Techniques: Knockdown, Western Blot, esiRNA, Quantitation Assay

Journal: iScience

Article Title: Role of human HSPE1 for OPA1 processing independent of HSPD1

doi: 10.1016/j.isci.2023.106067

Figure Lengend Snippet:

Article Snippet: The antibodies used were HSPD1 (1:1,000 dilution, 12165, Cell Signaling), DNAJA3 (1:1,000 dilution, 11088-1-AP; Proteintech), HSPA9 (1:5,000 dilution, 14887-1-AP; Proteintech), HSPA14 (1:1,000 dilution, MA5-32413, Invitrogen), OPA1 (1:1,000 dilution, 612607; BD Biosciences), GAPDH (1:1,000 dilution, MA5-15738; Invitrogen), TOM20 (1:2,000 dilution, sc-11415; Santa Cruz Biotechnology), OMA1 (1:500 dilution, sc-515788; Santa Cruz Biotechnology), MFN1/2 (1:1,000 dilution, ab57602; Abcam), PDH (1:1,000 dilution, ab110338; Abcam), TIM23 (1:5,000 dilution, 611223; BD), and GFP (1:5,000 dilution, a gift from Dr. Peter Devreotes).

Techniques: Recombinant, Modification, Saline, Protease Inhibitor, Plasmid Preparation, SYBR Green Assay, cDNA Synthesis, CRISPR, Cell Culture, Sequencing, esiRNA, Negative Control, Software