Journal: Nature Communications

Article Title: Prefusion-stabilized Hantaan virus glycoprotein nucleic acid vaccine elicits potent neutralizing antibody responses via germinal center activation

doi: 10.1038/s41467-026-70285-7

Figure Lengend Snippet: a Schematic of the prefusion-stabilized HTNV GP mRNA construct, with domains labeled: Gn (red), Gc (yellow), UTRs (black), Cap1 structure (5’ cap). The engineered cysteines for disulfide stabilization are marked by blue asterisks. b Representative negative-stain transmission electron microscopy images of GP-WT (left) and GP-C3 (right) mRNA-LNP particles. Scale bar, 100 nm. c In vitro expression validation. Huh7 cells were treated with GP-WT or GP-C3 mRNA-LNP or empty LNP, and protein expression was detected using an HTNV Gc-specific monoclonal antibody (3G1). Scale bar, 20 μm. d Immunization and challenge scheme. Female BALB/c mice (6–8 weeks old) received two i.m. doses of vaccine at a 3-week interval. Blood and spleens were collected at the indicated times for immune analysis. For the challenge, mice were inoculated with HTNV on day 28 and sacrificed on day 31. e , f Humoral immune responses post-immunization: HTNV GP-specific IgG titers ( e ) and HTNV-neutralizing antibody (NAb) titers ( f ) at days 14 and 28. The dashed line indicates the detection limit. g Cross-neutralizing antibody titers against Seoul virus (SEOV) in serum at day 28. h – o Cellular immune responses in splenocytes at day 28. Frequencies of IFN-γ– ( h ) and IL-2–secreting cells (i) (ELISpot), IFN-γ + CD4 + ( j ) and CD8 + T cells ( k ), IL-4–secreting cells ( l ) (ELISpot), IL-4 + CD4 + ( m ) and CD8 + T cells ( n ), and IL-10–secreting cells ( o ) (ELISpot) in response to HTNV-GP peptide stimulation. Data are presented as mean ± SEM. For ELIZA and ELISpot assays ( e – i , l , p), n = 5 mice per group. For ( b , c ), this experiment was independently repeated three times with similar results. For flow cytometry analyses ( j , k , m , n ), n = 3 mice per group. Statistical significance was determined by two-way ANOVA ( e , f ) or one-way ANOVA ( g – p ) with Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Exact P -values are provided in the Source Data file. Source data are provided as a Source Data file.

Article Snippet: Vero E6 (ATCC, CRL-1586), Huh7 (Procell, Wuhan, China; CL-0120), and HEK-293T (Clontech, now Takara Bio, 632180) cells were maintained in Dulbecco’s modified Eagle’s medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with l-glutamine, sodium pyruvate, and 10% fetal bovine serum (Sigma-Aldrich).

Techniques: Construct, Labeling, Staining, Transmission Assay, Electron Microscopy, In Vitro, Expressing, Biomarker Discovery, Virus, Enzyme-linked Immunospot, Flow Cytometry

Journal: MedComm

Article Title: Lycorine Derivative Inhibits SARS‐CoV‐2 Replication by Reducing −1 Programmed Ribosomal Frameshifting via Targeting ZAP

doi: 10.1002/mco2.70715

Figure Lengend Snippet: Lycorine derivatives exhibit potent antiviral activity against SARS‐CoV‐2 and its variants in vitro. (A) Dose‐dependent inhibition of SARS‐CoV‐2 original strain (GD108) infection by selected lycorine derivatives. Vero cells infected with SARS‐CoV‐2 at MOI of 0.05 were treated with serially diluted compounds for 48 h. Viral RNA copy numbers in culture media were quantified by RT‐qPCR (real‐time quantitative PCR). The dose‐inhibition curve for each compound is shown above the corresponding plot ( n = 3). Cytotoxicity assay of lycorine derivatives. Vero cells were treated with lycorine derivatives at gradient concentrations for 48 h. Cytotoxicity was assayed by CCK‐8 ( n = 3). (B) Vero cells were infected with different strains of SARS‐CoV‐2 (Alpha, Beta, Delta, and Omicron BA.1) at MOI of 0.05 and treated with serially diluted compounds ( 7 , 11 , 14 , and remdesivir) for 48 h. Viral RNA copy numbers in culture media were quantified by RT‐qPCR. The EC 50 value for each compound is shown above the corresponding plot ( n = 3). (C) IF of the inhibition of compound 7 on SARS‐CoV‐2 replication in Vero, Calu‐3, Huh‐7, and Caco‐2. At 48 h, the infected cells (MOI = 0.05) treated with different concentrations of compound 7 were fixed and analyzed by IF using the primary antibody against SARS‐CoV‐2 N protein (Green). Cell nuclei were stained with DAPI (blue). Scale bars = 100 µm. (D) WB analysis of the inhibition of compound 7 on SARS‐CoV‐2 replication in Vero, Calu‐3, Huh‐7, and Caco‐2. The cells were infected with SARS‐CoV‐2 GD108 at MOI of 0.05 and then treated with compound 7 . N protein expression was detected by WB at 48 h post‐infection ( n = 3).

Article Snippet: The human lung adenocarcinoma Calu‐3 cell line (Procell, CL‐0054, Wuhan, China), human hepatoma Huh‐7 cell line (Procell, CL‐0120), human colon Caco‐2 cell line (Procell, CL‐0050), human lung adenocarcinoma H1299 cell line (Procell, CL‐0165), and human embryonic kidney HEK293T cell line (Procell, CL‐0005) were cultured in standard medium at 37°C and 5% CO 2 .

Techniques: Activity Assay, In Vitro, Inhibition, Infection, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Cytotoxicity Assay, CCK-8 Assay, Staining, Expressing

Journal: MedComm

Article Title: Lycorine Derivative Inhibits SARS‐CoV‐2 Replication by Reducing −1 Programmed Ribosomal Frameshifting via Targeting ZAP

doi: 10.1002/mco2.70715

Figure Lengend Snippet: Compound 7 directly binds to ZAP and leads to a decrease in –1PRF. (A) Schematic representation of the dual‐luciferase frameshift reporter construct. The coding sequences for Renilla luciferase and Firefly luciferase were separated by the SARS‐CoV‐2 –1FSE sequence (13460‐13548). (B) Fold change in the Firefly/Renilla (F/R) luciferase ratio after treatment with the indicated drugs (lycorine at 10 µM, compound 7 at 10 µM, merafloxacin at 40 µM). Huh‐7 and H1299 cells transfected with the pHRF‐FSE (–1) luciferase reporter vector were treated with DMSO (Control) or the indicated drugs for 48 h ( n = 6 per group). (C) The frameshift Reporter mRNA containing a 3×FLAG‐tag followed by nucleotides 12686–14190 of the SARS‐CoV‐2 genome was translated in a RRL translation system in the presence of compound 7 or merafloxacin. The 3×FLAG‐tag was introduced at the N‐terminus to facilitate detection. WB analysis of the compound 7 effect on the –1PRF frameshift efficiency (FE) using anti‐flag antibody (Sigma, F3165). BC (blank control), NC (negative control). (D) Relative abundance of Nsp9, Nsp12, and Nsp15 in compound 7 (0, 2.5, and 5 µM) or Mer (merafloxacin, 40 µM)‐treated Vero cells after SARS‐CoV‐2 infection (MOI = 0.05). (E) The cellular target of compound 7 was identified using DARTS technology coupled with LC–MS/MS in H1299 cells. M, marker. (F) Venn diagram between the target proteins of compound 7 and the in vitro RNA antisense purification of the SARS‐CoV‐2 frameshift site from the literature. (G) ZAP protein stability was increased upon compound 7 (10 µM) treatment in H1299 cell lysates. (H) H1299 cells were transfected with the pcDNA3.1‐3×Flag‐Nsp12 plasmid and cultured for 48 h. Cells were then lysed and treated with 10 µM compound 7 (+) or DMSO (–). Recombinant Nsp12 (rNsp12) protein was detected by WB analysis, which was performed using anti‐Flag and anti‐Nsp12 antibodies for detection. (I) CETSA confirmed the binding of compound 7 (50 µM) to ZAP in 293T cells, with GAPDH serving as the internal control. (J) The binding of compound 7 to ZAP was depicted through BLI.

Article Snippet: The human lung adenocarcinoma Calu‐3 cell line (Procell, CL‐0054, Wuhan, China), human hepatoma Huh‐7 cell line (Procell, CL‐0120), human colon Caco‐2 cell line (Procell, CL‐0050), human lung adenocarcinoma H1299 cell line (Procell, CL‐0165), and human embryonic kidney HEK293T cell line (Procell, CL‐0005) were cultured in standard medium at 37°C and 5% CO 2 .

Techniques: Luciferase, Construct, Sequencing, Transfection, Plasmid Preparation, Control, Negative Control, Infection, Liquid Chromatography with Mass Spectroscopy, Marker, In Vitro, Purification, Cell Culture, Recombinant, Binding Assay

Journal: MedComm

Article Title: Lycorine Derivative Inhibits SARS‐CoV‐2 Replication by Reducing −1 Programmed Ribosomal Frameshifting via Targeting ZAP

doi: 10.1002/mco2.70715

Figure Lengend Snippet: Compound 7 exerts antiviral efficacy dependent upon ZAP‐S. (A) Effects of ZAP‐S knockdown on –1PRF followed by DMSO or compound 7 (10 µM) treatment in Huh‐7 and H1299 cells ( n = 3). (B) Compound 7 (10 µM) in combination with ZAP overexpression synergistically decreased –1PRF in H1299 and Huh‐7 cells ( n = 4); (C and D) Antiviral activity of compound 7 (2.5 µM) following ZAP‐S knockdown, or overexpression of ZAP‐S and mutant ZAP‐S. IF visualization of SARS‐CoV‐2 N protein (green) and cell nuclei (blue) in infected Huh‐7 cells at 48 h posttreatment (left). Scale bar: 100 µm. WB of N protein expression in Huh‐7 cells infected with SARS‐CoV‐2 (right).

Article Snippet: The human lung adenocarcinoma Calu‐3 cell line (Procell, CL‐0054, Wuhan, China), human hepatoma Huh‐7 cell line (Procell, CL‐0120), human colon Caco‐2 cell line (Procell, CL‐0050), human lung adenocarcinoma H1299 cell line (Procell, CL‐0165), and human embryonic kidney HEK293T cell line (Procell, CL‐0005) were cultured in standard medium at 37°C and 5% CO 2 .

Techniques: Knockdown, Over Expression, Activity Assay, Mutagenesis, Infection, Expressing

Journal: CytoJournal

Article Title: SOX2 promotes the progression of hepatocellular carcinoma by targeting FOXJ3 and activating the PI3K/AKT pathway to regulate autophagy and mitophagy

doi: 10.25259/Cytojournal_164_2025

Figure Lengend Snippet: SOX2 is highly expressed in hepatocellular carcinoma (HCC) cell lines. (a-c) qRT-PCR and Western blot analysis of SOX2 mRNA and protein expression levels in normal liver epithelial (THLE2) and HCC (Huh7) cell lines. (d-f) qRT-PCR and Western blot validation of transfection efficiency in Huh7 cells transfected with shRNA targeting SOX2 (shSOX2) or SOX2 overexpression plasmids. n = 6; ✶ ✶ ✶ P <0.001. SOX2: SRY-box transcription factor 2, qRT-PCR: Quantitative reverse transcription polymerase chain reaction. mRNA: messenger RNA; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; sh-NC: shRNA-negative control; Oe-NC: overexpression plasmid negative control.

Article Snippet: Human normal liver epithelial cells (THLE-2, CL-0833) and HCC cells (Huh7, CL-0120) were purchased from Procell (Wuhan, Hubei, China).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Biomarker Discovery, Transfection, shRNA, Over Expression, Reverse Transcription, Polymerase Chain Reaction, Negative Control, Plasmid Preparation

Journal: CytoJournal

Article Title: SOX2 promotes the progression of hepatocellular carcinoma by targeting FOXJ3 and activating the PI3K/AKT pathway to regulate autophagy and mitophagy

doi: 10.25259/Cytojournal_164_2025

Figure Lengend Snippet: SOX2 promotes the growth of HCC cells. (a and b) EdU staining assessing the effects of SOX2 overexpression or knockdown on Huh7 cell proliferation. (a) Magnification = ×100, Scale bar: 100 μm. (c-f) Transwell assays evaluating the effect of SOX2 on Huh7 cell motility. (c and e) Magnification = ×200, Scale bar: 50 μm. (g and h) Flow cytometry analysis of SOX2 effects on apoptosis in Huh7 cells (PI and Annexin V-FITC stain). n = 6; ✶ ✶ ✶ P <0.001. SOX2: SRY-box transcription factor 2, HCC: Hepatocellular carcinoma. sh-NC: shRNA-negative control; Oe-NC: Overexpression plasmid negative control; ECD-A: Energy Coupled Dye -Area; FITC-A: Fluorescein isothiocyanate -Area.

Article Snippet: Human normal liver epithelial cells (THLE-2, CL-0833) and HCC cells (Huh7, CL-0120) were purchased from Procell (Wuhan, Hubei, China).

Techniques: Staining, Over Expression, Knockdown, Flow Cytometry, shRNA, Negative Control, Plasmid Preparation

Journal: CytoJournal

Article Title: SOX2 promotes the progression of hepatocellular carcinoma by targeting FOXJ3 and activating the PI3K/AKT pathway to regulate autophagy and mitophagy

doi: 10.25259/Cytojournal_164_2025

Figure Lengend Snippet: SOX2 induces autophagy and mitophagy in HCC cells. (a-c) Western blot analysis of the effect of SOX2 on the expression levels of autophagy-related proteins LC3II, LC3I, and p62. (d) Transmission electron microscopic observation of autophagosome formation in HCC cells following SOX2 modulation. (e-g) Western blot analysis of the effect of SOX2 on mitophagy-related proteins PINK1, phosphorylated COX4 (p-COX4), and COX4. (h) Immunofluorescence analysis of the colocalization between mitochondria and lysosomes upon SOX2 expression. (i) Relative intensity of overlapping fluorescence signals of mitochondria and lysosomes. (d) and (h) magnification = ×200, Scale bar: 50 μm. n = 6; ✶ P <0.05, ✶ ✶ ✶ P <0.001. SOX2: SRY-box transcription factor 2, HCC: Hepatocellular carcinoma. GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; sh-NC: shRNA-negative control; Oe-NC: overexpression plasmid negative control.

Article Snippet: Human normal liver epithelial cells (THLE-2, CL-0833) and HCC cells (Huh7, CL-0120) were purchased from Procell (Wuhan, Hubei, China).

Techniques: Western Blot, Expressing, Transmission Assay, Immunofluorescence, Fluorescence, shRNA, Negative Control, Over Expression, Plasmid Preparation

Journal: CytoJournal

Article Title: SOX2 promotes the progression of hepatocellular carcinoma by targeting FOXJ3 and activating the PI3K/AKT pathway to regulate autophagy and mitophagy

doi: 10.25259/Cytojournal_164_2025

Figure Lengend Snippet: SOX2 regulates the expression of FOXJ3 in HCC. (a-c) qRT-PCR and Western blot analysis of FOXJ3 mRNA and protein expression levels in Huh7 cells following SOX2 knockdown and overexpression. n = 6; ✶ ✶ ✶ P <0.001. SOX2: SRY-box transcription factor 2, HCC: Hepatocellular carcinoma, FOXJ3: Forkhead box J3, qRT-PCR: Quantitative reverse transcription polymerase chain reaction.

Article Snippet: Human normal liver epithelial cells (THLE-2, CL-0833) and HCC cells (Huh7, CL-0120) were purchased from Procell (Wuhan, Hubei, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Over Expression, Reverse Transcription, Polymerase Chain Reaction

Journal: CytoJournal

Article Title: SOX2 promotes the progression of hepatocellular carcinoma by targeting FOXJ3 and activating the PI3K/AKT pathway to regulate autophagy and mitophagy

doi: 10.25259/Cytojournal_164_2025

Figure Lengend Snippet: SOX2 regulates autophagy and mitophagy in HCC cells by regulating FOXJ3. (a-c) Western blot analysis of the effects of SOX2 and FOXJ3 on the expression levels of autophagy-related proteins LC3II, LC3I, and p62. (d) Transmission electron microscopic observation of autophagosome formation in HCC cells under SOX2 and FOXJ3 modulation. Scale bar: 50 nm. (e-g) Western blot analysis of the effects of SOX2 and FOXJ3 on mitophagy-related proteins PINK1, phosphorylated COX4 (p-COX4), and COX4. (h and i) Immunofluorescence assay assessing the colocalization of mitochondria and lysosomes upon SOX2 and FOXJ3 expression. (h) Magnification = ×200, Scale bar: 50 μm. n = 6; ✶ ✶ ✶ P <0.001. SOX2: SRY-box transcription factor 2, HCC: Hepatocellular carcinoma, FOXJ3: Forkhead box J3.

Article Snippet: Human normal liver epithelial cells (THLE-2, CL-0833) and HCC cells (Huh7, CL-0120) were purchased from Procell (Wuhan, Hubei, China).

Techniques: Western Blot, Expressing, Transmission Assay, Immunofluorescence

Journal: Nature Communications

Article Title: A nuclease-mimetic platinum nanozyme induces concurrent DNA platination and oxidative cleavage to overcome cancer drug resistance

doi: 10.1038/s41467-022-35022-w

Figure Lengend Snippet: a CLSM images of intracellular localization of FITC-labelled NMPNs and FITC-labelled PNPs after incubated for 6 h under different pH conditions. Scale bar: 20 μm. Arrows indicate NMPNs in the cytoplasm after the escape from endosomes in cisplatin-resistant Huh7 cells. b CLSM images of intracellular localization of FITC-labelled NMPNs and FITC-labelled PNPs after incubated for 12 h under different pH conditions. Scale bar: 40 μm. Asterisks indicate NMPNs in the nucleus of cisplatin-resistant Huh7 cells. c Quantitative analysis of the colocalization between lysotracker and FITC-labelled NMPNs or PNPs. d Quantitative analysis of the colocalization between DAPI and FITC-labelled NMPNs or PNPs. e Bio-TEM images of cells after incubation for 12 h with NMPNs under different pH conditions. Arrowheads indicate NMPNs accumulated in the nucleus of Huh7 cells. Black asterisks indicate NMPNs in nucleus. Scale bar: 1 μm. f Schematic diagram of the NMPNs to target the tumour cell nucleus. All data are presented as means ± SEM, n = 3 independent experiments. Statistical significance was analyzed by one-way ANOVA with multiple comparisons test ( c , d ). Source data are provided as a Source Data file.

Article Snippet: L02 cells (CL-0111) and Huh7 (CL-0120) cells were obtained from iCell Bioscience Inc. (Shanghai, China).

Techniques: Incubation

Journal: Nature Communications

Article Title: A nuclease-mimetic platinum nanozyme induces concurrent DNA platination and oxidative cleavage to overcome cancer drug resistance

doi: 10.1038/s41467-022-35022-w

Figure Lengend Snippet: a The level of Pt-DNA adducts in the nucleus of cisplatin-resistant Huh7 cells after treatment with NMPNs at pH 6.5. Scale bar: 40 μm. Scale bar: 40 μm. b ROS levels in the nucleus of cisplatin-resistant Huh7 cells after treatment with PNPs or NMPNs at pH 6.5. Scale bar: 40 μm. c Immunofluorescence of the γ-H2AX in cisplatin-resistant Huh7 cells after different treatments at pH 6.5. Scale bar: 40 μm. d Immunofluorescence of the XPA and Pt-DNA adducts in cisplatin-resistant Huh7 cells after different treatments at pH 6.5. Scale bar: 40 μm. e Quantitative analysis of the colocalization between XPA and Pt-DNA adducts. f Immunofluorescence of the XPF and Pt-DNA adducts in cisplatin-resistant Huh7 cells after different treatments at pH 6.5. Scale bar: 40 μm. g Quantitative analysis of the colocalization between XPF and Pt-DNA adducts. h Western blot analysis of XPF expression in the nucleus of cisplatin-resistant Huh7 cells after different treatments. i Quantitative analysis of XPF expression in cisplatin-resistant Huh7 cells after different treatments at pH 6.5. j The schematic illustration of the mechanism underlying NMPNs to induce DNA platination and oxidative cleavage to combat Pt resistance in tumour cells. In comparison to Pt compounds and PNPs that cannot effectively generate ROS in the nucleus, NMPNs can readily accumulate in the nucleus by acidity-induced exposure of TAT peptides, and induce in situ ROS generation to induce DNA oxidative cleavage, thus destroying the DNA conformation required for NER. It hampers the recruitment of XPA and XPF and thus inhibiting NER pathway. All data are presented as means ± SEM, n = 3 independent experiments. Statistical significance was analyzed by one-way ANOVA with multiple comparisons test. Source data are provided as a Source Data file.

Article Snippet: L02 cells (CL-0111) and Huh7 (CL-0120) cells were obtained from iCell Bioscience Inc. (Shanghai, China).

Techniques: Immunofluorescence, Western Blot, Expressing, Comparison, In Situ

Journal: Nature Communications

Article Title: A nuclease-mimetic platinum nanozyme induces concurrent DNA platination and oxidative cleavage to overcome cancer drug resistance

doi: 10.1038/s41467-022-35022-w

Figure Lengend Snippet: a Immunofluorescence of the Pt-DNA adducts in cisplatin-resistant Huh7 cells after different treatments at pH 6.5. Scale bar: 40 μm. b Immunofluorescence of the Pt-DNA adducts in cisplatin-resistant Huh7 cells after treatments with NMPNs or NMPNs+NAC at different time points. Scale bar: 40 μm. c Quantitative analysis of Pt-DNA adducts in cisplatin-resistant Huh7 cells after treatments with NMPNs or NMPNs+NAC at different time points. n = 6 independent experiments. d Quantitative analysis of DNA damage of cisplatin-resistant Huh7 cells after treatment with cisplatin or NMPNs. Cisplatin: n = 46; NMPNs: n = 44. e Flow cytometry analysis of cell apoptosis after different treatments at pH 6.5. f and corresponding quantitative results. n = 5 independent experiments. g The inhibition effect of cisplatin and NMPNs on cisplatin-resistant Huh7 cells growth at different incubation times. h The cell viabilities of cisplatin-resistant Huh7 cells or si XPA -transfected cisplatin-resistant Huh7 cells after treatment with NMPNs, PNPs or cisplatin. n = 4 independent experiments. All the data are presented as means ± SEM, Statistical significance was analyzed by one-way ANOVA with multiple comparisons test. Source data are provided as a Source Data file.

Article Snippet: L02 cells (CL-0111) and Huh7 (CL-0120) cells were obtained from iCell Bioscience Inc. (Shanghai, China).

Techniques: Immunofluorescence, Flow Cytometry, Inhibition, Incubation, Transfection