Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: RP11-115N4.1 overexpression significantly decreased K562 cell proliferation. (A) EdU assay for cell proliferation in K562 cells with RP11-115N4.1 overexpression. The percentage of EdU positive cells was detected by flow cytometry. The picture scale was 100 μ m. (B) Cell cycle assay of K562 cells with RP11-115N4.1 overexpression. (C) Cell apoptosis analysis in K562 cells with RP11-115N4.1 overexpression. **** P < 0.0001. Each experiment was repeated three times and results are means ± SD.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Over Expression, EdU Assay, Flow Cytometry, Cell Cycle Assay

Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: RP11-115N4.1 promoted HSP70 transcription. (A) Volcano plot described that the differentially expressed mRNAs in K562 cells with RP11-115N4.1 overexpression. (B) GO analysis data described that the differentially expressed mRNAs in K562 cells with RP11-115N4.1 overexpression. (C, D) Validation of differentially expressed genes by qRT-PCR. * P < 0.05, ** P < 0.01. Each experiment was repeated three times and results are means ± SD.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Over Expression, Biomarker Discovery, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: RP11-115N4.1 promoted HSP70 expression by interacting with HNRNPH3. (A, B) RNA-pull-down assay to identify RP11-115N4.1 binding protein in K562 cells. The eluted proteins were separated by SDS-PAGE and subjected to silver staining. Antisense RNA to RP11-115N4.1 (AS) was used as a negative control. The red arrow indicates the band representing the RP11-115N4.1-specific binding protein identified by mass spectrometry as HNRNPH3. Western blot analysis confirmed that RP11-115N4.1 interacts with HNRNPH3. (C) RIP assay was performed using normal mouse IgG or the anti-HNRNPH3 antibody. GAPDH was used as the negative control. (D) Western blot of HSP70 and HNRNPH3 in K562 cells under different condition. β -Actin was used as an internal control. *** P < 0.001, **** P < 0.0001. Each experiment was repeated three times and results are means ± SD.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Expressing, Pull Down Assay, Binding Assay, SDS Page, Silver Staining, Negative Control, Mass Spectrometry, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: The supernatant of K562 cells overexpressed RP11-115N4.1 induced the inflammatory response of monocytes and inhibited the migration of trophoblast cells. (A) The level of HSP70 was determined by ELISA in K562 cell treated with RP11-115N4.1. (B) The level of HSP70 was determined by ELISA in the serum of URSA samples with high RP11-115N4.1 expression. (C) The levels of IL-6, IL-1β and TNF-α were determined by ELISA in human monocyte under different condition. LPS was used as a positive control (100 μg/mL). (D) Transwell assay in Swan 71 cell under different condition. The picture scale was 100 μ m. ** P < 0.01, *** P < 0.001, **** P < 0.0001. Each experiment was repeated three times and results are means ± SD.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Migration, Enzyme-linked Immunosorbent Assay, Expressing, Positive Control, Transwell Assay

Journal: Frontiers in Immunology

Article Title: Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion

doi: 10.3389/fimmu.2021.717785

Figure Lengend Snippet: Schematic diagram of role of RP11-115N4.1. RP11-115N4.1 bound to HNRNPH3 and increased the protein level of HSP70 in K562 cells. HSP70 released outside the cell induced the upregulation of the inflammatory factor IL-6, IL-1 β and TNF- α of monocytes and inhibited the migration of Swan 71. The black dashed line represents an unknown mechanism.

Article Snippet: K562 as chronic myelogenous leukemia cell line was purchased from Cell cook Biotech.

Techniques: Migration