Journal: Annals of Thoracic Medicine

Article Title: The diagnostic significance of signal peptide-complement C1r/C1s, Uegf, and Bmp1-epidermal growth factor domain-containing protein-1 levels in pulmonary embolism

doi: 10.4103/1817-1737.191876

Figure Lengend Snippet: SCUBE1 levels in PE, non-PE, and control groups. Horizontal lines represent the median of SCUBE1 levels for PE, non-PE, and control groups as 0.90, 0.38, and 0.47 ng/mL, respectively. SCUBE1: Signal peptide-complement C1r/C1s, Uegf, and Bmp1-epidermal growth factor domain-containing protein 1

Article Snippet: Human SCUBE1 ELISA kit (Elabscience Biotechnology Co., Ltd., China, Catalog no: E-EL-H5405, Lot: AK0015NOV30024) was used with BIOTEK semiautomatic ELISA reader.

Techniques: Control

Journal: Annals of Thoracic Medicine

Article Title: The diagnostic significance of signal peptide-complement C1r/C1s, Uegf, and Bmp1-epidermal growth factor domain-containing protein-1 levels in pulmonary embolism

doi: 10.4103/1817-1737.191876

Figure Lengend Snippet: Receiver operating characteristics curve analysis of plasma SCUBE1 values in pulmonary embolism patients. The optimum diagnostic cutoff point of SCUBE1 was 0.49 ng/mL; the area under the curve was 0.791 (95% confidence interval: 0.945–1.000); the sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 64%, 56%, and 100%, respectively. SCUBE1: Signal peptide-complement C1r/C1s, Uegf, and Bmp1-epidermal growth factor domain-containing protein 1

Article Snippet: Human SCUBE1 ELISA kit (Elabscience Biotechnology Co., Ltd., China, Catalog no: E-EL-H5405, Lot: AK0015NOV30024) was used with BIOTEK semiautomatic ELISA reader.

Techniques: Clinical Proteomics, Diagnostic Assay

Journal: Cell Death & Disease

Article Title: Histocompatibility Minor 13 (HM13), targeted by miR-760, exerts oncogenic role in breast cancer by suppressing autophagy and activating PI3K-AKT-mTOR pathway

doi: 10.1038/s41419-022-05154-4

Figure Lengend Snippet: A , B The expression of HM13 according to TCGA based on sample types ( A ) or subclasses ( B ) in breast cancer. Transcriptome profiling was obtained from TCGA. Based on PAM50 signature, we processed the data and showed the expression level of HM13 in breast cancer subclasses. P -values smaller than 0.05. C , D The correlation between HM13 expression and prognosis of the breast cancer patients was presented by Kaplan-Meier survival analysis. E , F qRT-PCR was used to evaluate the mRNA expression levels of HM13 in breast cancer tissues ( E ) and cell lines ( F ), respectively. G , H Protein expression of HM13 in pairs of tissues ( G ) and cell lines ( H ) of breast cancer. Data were shown as mean ± SD, * p < 0.05, ** p < 0.01.

Article Snippet: After the incubation of the primary antibody against HM13 (1:1000, Proteintech, USA, 20416-1-AP), PERK (1:1000, Cell Signaling Technology, USA, 5683 T), CHOP (1:1000, Cell Signaling Technology, USA, 2895 T), LC3B (1:1000, Cell Signaling Technology, USA, 3868 T), p62 (1:1000, Cell Signaling Technology, USA, 16177 S), BNIP3 (1:1000, Cell Signaling Technology, USA, 44060 S), p-PI3K (1:1000, Cell Signaling Technology, USA, 17366 S), PI3K (1:1000, Cell Signaling Technology, USA, 4249 T), p-AKT (1:1000, Cell Signaling Technology, USA, 4060 T), AKT (1:1000, Cell Signaling Technology, USA, 4691 T), p-mTOR (1:1000, Cell Signaling Technology, USA, 5536 T), mTOR (1:1000, Cell Signaling Technology, USA, 2983 T), GAPDH (1:1000, Beyotime, China, AF1186) and secondary antibodies, the relative protein expression levels derived from the bands were detected by an ECL detection system.

Techniques: Expressing, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: Histocompatibility Minor 13 (HM13), targeted by miR-760, exerts oncogenic role in breast cancer by suppressing autophagy and activating PI3K-AKT-mTOR pathway

doi: 10.1038/s41419-022-05154-4

Figure Lengend Snippet: Expression of HM13 in human breast cancer patients’ tissues according to patients’ clinicopathological.

Article Snippet: After the incubation of the primary antibody against HM13 (1:1000, Proteintech, USA, 20416-1-AP), PERK (1:1000, Cell Signaling Technology, USA, 5683 T), CHOP (1:1000, Cell Signaling Technology, USA, 2895 T), LC3B (1:1000, Cell Signaling Technology, USA, 3868 T), p62 (1:1000, Cell Signaling Technology, USA, 16177 S), BNIP3 (1:1000, Cell Signaling Technology, USA, 44060 S), p-PI3K (1:1000, Cell Signaling Technology, USA, 17366 S), PI3K (1:1000, Cell Signaling Technology, USA, 4249 T), p-AKT (1:1000, Cell Signaling Technology, USA, 4060 T), AKT (1:1000, Cell Signaling Technology, USA, 4691 T), p-mTOR (1:1000, Cell Signaling Technology, USA, 5536 T), mTOR (1:1000, Cell Signaling Technology, USA, 2983 T), GAPDH (1:1000, Beyotime, China, AF1186) and secondary antibodies, the relative protein expression levels derived from the bands were detected by an ECL detection system.

Techniques: Expressing

Journal: Cell Death & Disease

Article Title: Histocompatibility Minor 13 (HM13), targeted by miR-760, exerts oncogenic role in breast cancer by suppressing autophagy and activating PI3K-AKT-mTOR pathway

doi: 10.1038/s41419-022-05154-4

Figure Lengend Snippet: A , B The transfection efficiencies of si-HM13-1 and si-HM13-2 were evaluated by qRT-PCR ( A ) and western blot ( B ) in SUM1315 and ZR-75-1 cell lines. C The CCK-8 assays were performed to measure the cell viability of SUM1315 (left) and ZR-75-1 (right) cell lines transfected with siRNAs. D Representative results of the colony formation showed the cell proliferation in SUM1315 and ZR-75-1 cell lines after downregulation of HM13. E , F EdU assays were conducted to compare the growth rates in SUM1315 ( E ) and ZR-75-1 ( F ) cell lines between experimental groups (si-HM13-1 and si-HM13-2) and control group (si-NC). DAPI was indicated by blue, EdU was indicated by red. Scale bars, 50 μm. G Images of xenograft tumors from nude mice classified into HM13 knockdown (sh-HM13) group and control (sh-NC) group ( n = 6). H , I Average tumor volume ( H ) and tumor weight ( I ) of breast cancer in knockdown of HM13 (sh-HM13) and control (sh-NC) group were shown by tumor growth curves. J IHC staining of breast cancer tissues from tumor subcutaneous mice model was aimed to determine the positive rates of HM13 and Ki-67 in knockdown of HM13 (sh-HM13) and control (sh-NC) group. Scale bars, 100 µm. Data were shown as mean ± SD, (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: After the incubation of the primary antibody against HM13 (1:1000, Proteintech, USA, 20416-1-AP), PERK (1:1000, Cell Signaling Technology, USA, 5683 T), CHOP (1:1000, Cell Signaling Technology, USA, 2895 T), LC3B (1:1000, Cell Signaling Technology, USA, 3868 T), p62 (1:1000, Cell Signaling Technology, USA, 16177 S), BNIP3 (1:1000, Cell Signaling Technology, USA, 44060 S), p-PI3K (1:1000, Cell Signaling Technology, USA, 17366 S), PI3K (1:1000, Cell Signaling Technology, USA, 4249 T), p-AKT (1:1000, Cell Signaling Technology, USA, 4060 T), AKT (1:1000, Cell Signaling Technology, USA, 4691 T), p-mTOR (1:1000, Cell Signaling Technology, USA, 5536 T), mTOR (1:1000, Cell Signaling Technology, USA, 2983 T), GAPDH (1:1000, Beyotime, China, AF1186) and secondary antibodies, the relative protein expression levels derived from the bands were detected by an ECL detection system.

Techniques: Transfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Control, Knockdown, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: Histocompatibility Minor 13 (HM13), targeted by miR-760, exerts oncogenic role in breast cancer by suppressing autophagy and activating PI3K-AKT-mTOR pathway

doi: 10.1038/s41419-022-05154-4

Figure Lengend Snippet: A SUM1315 (left) and ZR-75-1 (right) cell lines were transfected with siRNAs (si-HM13-1 and si-HM13-2) or negative control (si-NC) and relative mRNA expression of ER stress-relating genes was analyzed by qRT–PCR. B Western blot was performed to detect the protein levels of PERK and CHOP in breast cancer cell lines in HM13 knockdown groups and control group. C , D SUM1315 ( C ) and ZR-75-1 ( D ) were stably transfected with mCherry-EGFP-LC3B lentivirus and then treated with siRNAs or si-NC, respectively. The subcellular localization of autophagosomes and autolysosomes were illustrated by fluorescent confocal microscopy and the bars graphs showed the specific quantities of the fluorescent puncta data. The yellow puncta indicated autophagosomes, and the red puncta indicated autolysosomes (the fusion of autophagosomes with lysosomes) in the merged figures. The data were presented as the mean ± SD of three experiments. Scale bars, 10 µm. E Representative transmission electron micrographs demonstrated the ultrastructure of breast cancer cells. Double membrane autophagosomes were counted in randomly selected >100 cells. Arrow indicates the autophagosomes distributing in the cytoplasm. Scale bars, 2 µm and 200 nm. F The protein expressions of LC3B-II, LC3B-I, and p62 in SUM1315 and ZR-75-1 respectively transfected with siRNAs or negative control were determined by western blot. Data were shown as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: After the incubation of the primary antibody against HM13 (1:1000, Proteintech, USA, 20416-1-AP), PERK (1:1000, Cell Signaling Technology, USA, 5683 T), CHOP (1:1000, Cell Signaling Technology, USA, 2895 T), LC3B (1:1000, Cell Signaling Technology, USA, 3868 T), p62 (1:1000, Cell Signaling Technology, USA, 16177 S), BNIP3 (1:1000, Cell Signaling Technology, USA, 44060 S), p-PI3K (1:1000, Cell Signaling Technology, USA, 17366 S), PI3K (1:1000, Cell Signaling Technology, USA, 4249 T), p-AKT (1:1000, Cell Signaling Technology, USA, 4060 T), AKT (1:1000, Cell Signaling Technology, USA, 4691 T), p-mTOR (1:1000, Cell Signaling Technology, USA, 5536 T), mTOR (1:1000, Cell Signaling Technology, USA, 2983 T), GAPDH (1:1000, Beyotime, China, AF1186) and secondary antibodies, the relative protein expression levels derived from the bands were detected by an ECL detection system.

Techniques: Transfection, Negative Control, Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Control, Stable Transfection, Confocal Microscopy, Transmission Assay, Membrane

Journal: Cell Death & Disease

Article Title: Histocompatibility Minor 13 (HM13), targeted by miR-760, exerts oncogenic role in breast cancer by suppressing autophagy and activating PI3K-AKT-mTOR pathway

doi: 10.1038/s41419-022-05154-4

Figure Lengend Snippet: A In SUM1315 and ZR-75-1 cell lines transfected with negative control (si-NC), siRNA (si-HM13), siRNA and DMSO (si+DMSO), siRNA and CQ (si+CQ), the protein levels of LC3B-II/I and p62 were analyzed by western blot, respectively. B , C Cell proliferation abilities were determined by CCK8 ( B ) and colony formation assays ( C ). D Representative images of the transwell assays using SUM1315 an ZR-75-1 cell lines transfected with negative control (si-NC), siRNA (si-HM13), siRNA and DMSO (si + DMSO), siRNA and CQ (si + CQ). Scale bars, 50 µm. Data were shown as mean ± SD, * p < 0.05, ** p < 0.01.

Article Snippet: After the incubation of the primary antibody against HM13 (1:1000, Proteintech, USA, 20416-1-AP), PERK (1:1000, Cell Signaling Technology, USA, 5683 T), CHOP (1:1000, Cell Signaling Technology, USA, 2895 T), LC3B (1:1000, Cell Signaling Technology, USA, 3868 T), p62 (1:1000, Cell Signaling Technology, USA, 16177 S), BNIP3 (1:1000, Cell Signaling Technology, USA, 44060 S), p-PI3K (1:1000, Cell Signaling Technology, USA, 17366 S), PI3K (1:1000, Cell Signaling Technology, USA, 4249 T), p-AKT (1:1000, Cell Signaling Technology, USA, 4060 T), AKT (1:1000, Cell Signaling Technology, USA, 4691 T), p-mTOR (1:1000, Cell Signaling Technology, USA, 5536 T), mTOR (1:1000, Cell Signaling Technology, USA, 2983 T), GAPDH (1:1000, Beyotime, China, AF1186) and secondary antibodies, the relative protein expression levels derived from the bands were detected by an ECL detection system.

Techniques: Transfection, Negative Control, Western Blot

Journal: Cell Death & Disease

Article Title: Histocompatibility Minor 13 (HM13), targeted by miR-760, exerts oncogenic role in breast cancer by suppressing autophagy and activating PI3K-AKT-mTOR pathway

doi: 10.1038/s41419-022-05154-4

Figure Lengend Snippet: A Western blot assessment of PI3K, p-PI3K, AKT, p-AKT, mTOR, p-mTOR protein expression was performed in SUM1315 and ZR-75-1 cell lines with siRNAs (si-HM13-1 and si-HM13-2) or negative control (si-NC). B , D The effects of IGF-1 on cell proliferation were detected by CCK-8 ( B ) and EdU ( C , D ) assays. SUM1315 and ZR-75-1 cell lines were transfected with negative control (si-NC), siRNA (si-HM13) or siRNA and IGF-1 (si+IGF-1) before harvesting. Scale bars, 50 µm. E , F The wound healing assays were performed to determine the migration abilities of SUM1315 ( E ) and ZR-75-1 ( F ) transfected with negative control (si-NC), siRNA (si-HM13) or siRNA and IGF-1 (si+IGF-1). Scale bars, 100 µm. Data were shown as mean ± SD, * p < 0.05, ** p < 0.01.

Article Snippet: After the incubation of the primary antibody against HM13 (1:1000, Proteintech, USA, 20416-1-AP), PERK (1:1000, Cell Signaling Technology, USA, 5683 T), CHOP (1:1000, Cell Signaling Technology, USA, 2895 T), LC3B (1:1000, Cell Signaling Technology, USA, 3868 T), p62 (1:1000, Cell Signaling Technology, USA, 16177 S), BNIP3 (1:1000, Cell Signaling Technology, USA, 44060 S), p-PI3K (1:1000, Cell Signaling Technology, USA, 17366 S), PI3K (1:1000, Cell Signaling Technology, USA, 4249 T), p-AKT (1:1000, Cell Signaling Technology, USA, 4060 T), AKT (1:1000, Cell Signaling Technology, USA, 4691 T), p-mTOR (1:1000, Cell Signaling Technology, USA, 5536 T), mTOR (1:1000, Cell Signaling Technology, USA, 2983 T), GAPDH (1:1000, Beyotime, China, AF1186) and secondary antibodies, the relative protein expression levels derived from the bands were detected by an ECL detection system.

Techniques: Western Blot, Expressing, Negative Control, CCK-8 Assay, Transfection, Migration

Journal: Cell Death & Disease

Article Title: Histocompatibility Minor 13 (HM13), targeted by miR-760, exerts oncogenic role in breast cancer by suppressing autophagy and activating PI3K-AKT-mTOR pathway

doi: 10.1038/s41419-022-05154-4

Figure Lengend Snippet: A The potential candidate miRNAs targeting HM13 were screened and presented by the Venn-diagram. B The relative mRNA expression of HM13 was analyzed by qRT-PCR. miR-6822-3p mimics (miR-6822-3p), miR-6747-3p mimics (miR-6747-3p), miR-149-5p mimics (miR-149-5p) or miR-760 mimics (miR-760) was transfected into breast cancer cell lines for 48 h before harvesting, respectively. C The mRNA expression of HM13 in SUM1315 (left) and ZR-75-1 (right) cell lines after transfected with miR-760 inhibitor (inh-760) and inhibitor control (inh-NC) was determined by qRT-PCR. D Relative protein expression of HM13 from four groups containing miR-760 inhibitor (inh-760) group, miR-760 mimics (miR-760) group and their corresponding control groups (inh-NC and miR-NC) was evaluated through western blot. E Schematic diagram of the regions in the 3′-UTR of HM13 mRNA. F Relative luciferase activities of wild type and mutant HM13 reporter plasmid in SUM1315 (left) and ZR-75-1 (right) cell lines transfected with miR-760 mimics (miR-760) or mimics control (miR-NC). Data were shown as mean ± SD, * p < 0.05, ** p < 0.01.

Article Snippet: After the incubation of the primary antibody against HM13 (1:1000, Proteintech, USA, 20416-1-AP), PERK (1:1000, Cell Signaling Technology, USA, 5683 T), CHOP (1:1000, Cell Signaling Technology, USA, 2895 T), LC3B (1:1000, Cell Signaling Technology, USA, 3868 T), p62 (1:1000, Cell Signaling Technology, USA, 16177 S), BNIP3 (1:1000, Cell Signaling Technology, USA, 44060 S), p-PI3K (1:1000, Cell Signaling Technology, USA, 17366 S), PI3K (1:1000, Cell Signaling Technology, USA, 4249 T), p-AKT (1:1000, Cell Signaling Technology, USA, 4060 T), AKT (1:1000, Cell Signaling Technology, USA, 4691 T), p-mTOR (1:1000, Cell Signaling Technology, USA, 5536 T), mTOR (1:1000, Cell Signaling Technology, USA, 2983 T), GAPDH (1:1000, Beyotime, China, AF1186) and secondary antibodies, the relative protein expression levels derived from the bands were detected by an ECL detection system.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, Western Blot, Luciferase, Mutagenesis, Plasmid Preparation

Journal: Cell Death & Disease

Article Title: Histocompatibility Minor 13 (HM13), targeted by miR-760, exerts oncogenic role in breast cancer by suppressing autophagy and activating PI3K-AKT-mTOR pathway

doi: 10.1038/s41419-022-05154-4

Figure Lengend Snippet: A , B Inhibitor control (inh-NC), miR-760 inhibitor (inh-760), miR-760 inhibitor and negative control (inh-760+si-NC) or miR-760 inhibitor and siRNA (inh-760+si-HM13) was transfected into SUM1315 and ZR-75-1 cell lines, respectively. The cell proliferation abilities were evaluated via CCK-8 ( A ), as well as colony formation assay ( B ). C , D SUM1315 and ZR-75-1 cell lines were divided into four groups, inhibitor control (inh-NC) group, miR-760 inhibitor (inh-760) group, miR-760 inhibitor and negative control (inh-760+si-NC) group or miR-760 inhibitor and siRNA (inh-760+si-HM13) group. The metastasis abilities were detected by the wound healing assays ( C ) and the transwell assays ( D ). E Western blot was aimed to evaluate the protein expression of genes related to autophagy and PI3K-AKT-mTOR signaling in rescue experiments. Data were shown as mean ± SD, * p < 0.01, ** p < 0.01.

Article Snippet: After the incubation of the primary antibody against HM13 (1:1000, Proteintech, USA, 20416-1-AP), PERK (1:1000, Cell Signaling Technology, USA, 5683 T), CHOP (1:1000, Cell Signaling Technology, USA, 2895 T), LC3B (1:1000, Cell Signaling Technology, USA, 3868 T), p62 (1:1000, Cell Signaling Technology, USA, 16177 S), BNIP3 (1:1000, Cell Signaling Technology, USA, 44060 S), p-PI3K (1:1000, Cell Signaling Technology, USA, 17366 S), PI3K (1:1000, Cell Signaling Technology, USA, 4249 T), p-AKT (1:1000, Cell Signaling Technology, USA, 4060 T), AKT (1:1000, Cell Signaling Technology, USA, 4691 T), p-mTOR (1:1000, Cell Signaling Technology, USA, 5536 T), mTOR (1:1000, Cell Signaling Technology, USA, 2983 T), GAPDH (1:1000, Beyotime, China, AF1186) and secondary antibodies, the relative protein expression levels derived from the bands were detected by an ECL detection system.

Techniques: Control, Negative Control, Transfection, CCK-8 Assay, Colony Assay, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: Histocompatibility Minor 13 (HM13), targeted by miR-760, exerts oncogenic role in breast cancer by suppressing autophagy and activating PI3K-AKT-mTOR pathway

doi: 10.1038/s41419-022-05154-4

Figure Lengend Snippet: Downregulated HM13 could activate autophagy by inducing ER-stress, while HM13 upregulation could facilitate PI3K-AKT-mTOR pathway in breast cancer. Moreover, miR-760 negatively targets HM13 by binding to 3′-UTR of its mRNA. Thus, HM13 plays an oncogenic role in breast cancer progression.

Article Snippet: After the incubation of the primary antibody against HM13 (1:1000, Proteintech, USA, 20416-1-AP), PERK (1:1000, Cell Signaling Technology, USA, 5683 T), CHOP (1:1000, Cell Signaling Technology, USA, 2895 T), LC3B (1:1000, Cell Signaling Technology, USA, 3868 T), p62 (1:1000, Cell Signaling Technology, USA, 16177 S), BNIP3 (1:1000, Cell Signaling Technology, USA, 44060 S), p-PI3K (1:1000, Cell Signaling Technology, USA, 17366 S), PI3K (1:1000, Cell Signaling Technology, USA, 4249 T), p-AKT (1:1000, Cell Signaling Technology, USA, 4060 T), AKT (1:1000, Cell Signaling Technology, USA, 4691 T), p-mTOR (1:1000, Cell Signaling Technology, USA, 5536 T), mTOR (1:1000, Cell Signaling Technology, USA, 2983 T), GAPDH (1:1000, Beyotime, China, AF1186) and secondary antibodies, the relative protein expression levels derived from the bands were detected by an ECL detection system.

Techniques: Binding Assay