Journal: American Journal of Cancer Research

Article Title: PRDM16 from hepatic stellate cells-derived extracellular vesicles promotes hepatocellular carcinoma progression

doi:

Figure Lengend Snippet: EVs-derived PRDM16 regulates the Notch signaling pathway in HCC cells. A. The KEGG enrichment analysis of genes positively correlated with PRDM16 in HCC. B. Correlation between NOTCH1 expression and PRDM16 expression in HCC. C. The schematic for the Notch signaling pathway. D. The effect of EVs treatment on Notch signaling pathway activity in HCC cells was measured using luciferase reporter assays. E. Effect of EVs treatment on the expression of NOTCH1 and the downstream factors HES1 and HES5 in HCC cells by RT-qPCR. F. The enrichment ability of PRDM16 on the NOTCH1 promoter was measured using ChIP-qPCR. G. The effect of PRDM16 carried by EVs on the transcriptional activity of the NOTCH1 promoter in HCC cells was measured using luciferase reporter assays. The results represent means ± SD, *P < 0.05 vs PBS; #P < 0.05 vs EVs-NC; &P < 0.05 vs IgG. All experiments were repeated at least three times, two-way ANOVA.

Article Snippet: The sections were sealed at room temperature for 1 h in 10% normal goat serum, followed by incubation with primary antibodies to PRDM16 (CSB-PA872534LA01HU, Cusabio Biotech, Newark, DE, USA), NOTCH1 (1:500, 10062-2-AP, ProteinTech Group), HES5 (1:200, 22666-1-AP, ProteinTech Group), and HES1 (1:1000, #11988, Cell Signaling Technologies) overnight at 4°C.

Techniques: Derivative Assay, Expressing, Activity Assay, Luciferase, Quantitative RT-PCR, ChIP-qPCR

Journal: American Journal of Cancer Research

Article Title: PRDM16 from hepatic stellate cells-derived extracellular vesicles promotes hepatocellular carcinoma progression

doi:

Figure Lengend Snippet: LX-2-derived EVs affect HCC growth in vivo via PRDM16/Notch signaling. (A) The volume of xenografts in mice. (B) Weight of harvested xenografts in mice. Immunohistochemical detection of PRDM16 (C), NOTCH1 (D), HES1 (E), and HES5 (F) expression in xenograft tumors. The results represent means ± SD (n = 8), *P < 0.05 vs PBS; #P < 0.05 vs EVs-NC; &P < 0.05 vs EVs-NC + DMSO. One-way or two-way ANOVA.

Article Snippet: The sections were sealed at room temperature for 1 h in 10% normal goat serum, followed by incubation with primary antibodies to PRDM16 (CSB-PA872534LA01HU, Cusabio Biotech, Newark, DE, USA), NOTCH1 (1:500, 10062-2-AP, ProteinTech Group), HES5 (1:200, 22666-1-AP, ProteinTech Group), and HES1 (1:1000, #11988, Cell Signaling Technologies) overnight at 4°C.

Techniques: Derivative Assay, In Vivo, Immunohistochemical staining, Expressing

Journal: bioRxiv

Article Title: A virus-induced circular RNA maintains latent infection of Kaposi sarcoma herpesvirus

doi: 10.1101/2022.07.18.500467

Figure Lengend Snippet: (A) Diagram of the RNA-pulldown assay is shown. Total RNA was incubated with biotinylated DNA oligos that is specific to circ_0001400 and circRNA-RNA complexes are purified using streptavidin beads. Next generation sequencing (NGS) was performed to determine their identities. (B) A volcano plot of transcripts identified by the circ_0001400 RNA pulldown assay in 293T cells. Mapped reads were median-scaled and enrichments were calculated by voom (limma package). ZKSCAN1 and RELL1 are partially included in the circ_0001400 expression plasmid vectors and thus serve as positive control of pulldown. Transcripts that were confirmed in are depicted as orange rectangles. n=3. (C) RT-qPCR of enriched transcripts after RNA-pulldown. circ_0001400 was ectopically expressed in 293T cells for 24 hours and RNA-pulldowns were performed (n=3). Enrichment of circ_0001400 and candidate mRNAs (listed in Table S5) are shown. (D) TTI1 transcript levels after manipulation of circ_0001400 transcript levels. circ_0001400 was ectopically expressed (n=6) or depleted by siRNAs (n=3) in 293T cells for 48 hours. Total RNAs were extracted followed by RT-qPCR. (E) Proposed model of circ_0001400’s regulation of PI3K/AKT/mTOR pathway. Significances were calculated with paired-t tests. *:p-value < 0.05.

Article Snippet: For primary endothelial cells, lentiviral vectors were used. pcDNA3.1-has_circ_0001400 and pcDNA3.1(+) ZKSCAN1 MCS-WT Split GFP + Sense IRES (Addgene plasmid # 69909) were digested with BamHI and XhoI (NEB) and cloned into pLV-mCHerry:T2A:Bsd-CMV plasmid (VectorBuilder).

Techniques: Incubation, Purification, Next-Generation Sequencing, Expressing, Plasmid Preparation, Positive Control, Quantitative RT-PCR

Journal: Pathology International

Article Title: FHL1: A novel diagnostic marker for papillary thyroid carcinoma

doi: 10.1111/pin.13467

Figure Lengend Snippet: Clinical, and histopathological characteristics of patients with papillary thyroid carcinoma (classical subtype).

Article Snippet: Slides were immunostained with monoclonal antibodies against the BCL2 antigen (clone SP66, Maixin), CD117/C‐kit antigen (clone YR145, Maixin), and with monoclonal antibodies against the FHL1 (10991‐1‐AP, Proteintech Group, Inc.), PPARGC1A (Boster, China), and TLE1 antigen (Maixin).

Techniques: Expressing

Journal: Pathology International

Article Title: FHL1: A novel diagnostic marker for papillary thyroid carcinoma

doi: 10.1111/pin.13467

Figure Lengend Snippet: The expression of FHL1, BCL2, TLE1, KIT, PPARGC1A and GHR in papillary thyroid carcinoma (PTC) and the control group. FHL1 showed strong positivity in the nuclei and to a lesser extent in the cytoplasm of normal thyroid tissues adjacent to the tumour (the control group), but is not present in the PTC tissues. ( p < 0.01). There was no significant difference in the expression of BCL2 between PTC and normal thyroid tissues ( p > 0.05). The results showed that TLE1, KIT, PPARGC1A, and GHR were absent in both tumor and normal tissues. (magnification ×200). FHL1, four and a half LIM domains 1.

Article Snippet: Slides were immunostained with monoclonal antibodies against the BCL2 antigen (clone SP66, Maixin), CD117/C‐kit antigen (clone YR145, Maixin), and with monoclonal antibodies against the FHL1 (10991‐1‐AP, Proteintech Group, Inc.), PPARGC1A (Boster, China), and TLE1 antigen (Maixin).

Techniques: Expressing, Control