sP10

The binding of sP10 and NPM1 was analyzed using Surface Plasmon Resonance (SPR) technology, which demonstrated that NPM had a strong affinity for sP10, and the equilibrium dissociation constant KD was found to be 9.216E-9 (Fig. 1G).

In vitro experiments confirmed that the IC50 of sP10-LNP on hepatoma cells decreased from 18.71 µM to 9.568 µM (with a 49% increase in activity) (Fig. 7E).

In vitro experiments confirmed that the IC50 of sP10-LNP on hepatoma cells decreased from 18.71 µM to 9.568 µM (with a 49% increase in activity) (Fig. 7E).

The proliferation curve demonstrated that both intracellular expression of peptide aptamers (P10 and P177) and incubation of synthetic peptides (sP10 and sP177) significantly decreased cell proliferation (Fig. 2A).

Our findings revealed that P10 and sP10 inhibited the proliferation of SK-Hep1 cells and Hepa1-6 cells, respectively, as demonstrated by the CCK-8 proliferation assay (Fig. 3A).

Our findings revealed that P10 and sP10 inhibited the proliferation of SK-Hep1 cells and Hepa1-6 cells, respectively, as demonstrated by the CCK-8 proliferation assay (Fig. 3A).

Apoptosis assays demonstrated that sP10 and sP177 promoted apoptosis in HCC cells (SK-Hep1, Hepa1-6) and dose-dependently in LX-2 cells (Fig. 4F, Fig. S6).

Apoptosis assays demonstrated that sP10 and sP177 promoted apoptosis in HCC cells (SK-Hep1, Hepa1-6) and dose-dependently in LX-2 cells (Fig. 4F, Fig. S6).

while showing no significant effects on AML12 normal hepatocytes (Fig. 4F).

Our findings revealed that sP10 significantly reduced the size and weight of transplanted tumors (Fig. 3F).

Further analysis of mouse lung surface metastases (Supplementary Fig. S4G) and IHC staining of lung tissue sections (Fig. 3H) revealed that sP10 significantly decreased the number of lung metastases.

Fluorescence analysis revealed that sP10 significantly reduced the intensity and range of fluorescence of transplanted tumors in living mice, indicating that sP10 inhibited tumor growth in mice (Fig. 3G).

Lung fluorescence of BALB/c-nu mice showed that sP10 significantly inhibited lung metastasis (Supplementary Fig. S4J).

The results of Sirius red staining, IHC analysis of liver tissues (as shown in Fig. 2E), and Western blotting (Fig. 2F) indicated that both sP10 and sP177 effectively decreased collagen synthesis and α-SMA expression in fibrotic liver tissue.

These results indicate that sP10 did not cause significant pathological changes in the structure and function of the major organs of mice, suggesting that sP10 has good safety (Fig. 8).

Confocal microscopy showed increased autophagosomes (yellow puncta) and autolysosomes (red puncta) after 24 h (Fig. 5E and F), confirming enhanced autophagic flux.

Confocal microscopy showed increased autophagosomes (yellow puncta) and autolysosomes (red puncta) after 24 h (Fig. 5E and F), confirming enhanced autophagic flux.

Flow cytometry analysis revealed that sP10 increased the proportion of CD80-positive and CD86-positive M1 macrophages derived from THP-1 cells (Fig. 6C).

Flow cytometry analysis revealed that sP10 increased the proportion of CD80-positive and CD86-positive M1 macrophages derived from BMDM cells (Fig. 6D).

Quantitative PCR demonstrated marked suppression of M2 markers (CD206, Arg-1, IL-10) (Fig. 6B).

WB analysis revealed significant downregulation of TGFβ1, α-SMA, and phosphorylated Smad2 (p-Smad2) (Fig. 6A).

sP10 significantly reduced cytoplasmic translocation of NPM1 in HCC cell lines (SK-Hep1, Hepa1-6, LM3) but not in normal hepatocytes AML12 (Fig. 4E).

sP10 significantly reduced cytoplasmic translocation of NPM1 in HCC cell lines (SK-Hep1, Hepa1-6, LM3) but not in normal hepatocytes AML12 (Fig. 4E).

sP10 significantly reduced cytoplasmic translocation of NPM1 in HCC cell lines (SK-Hep1, Hepa1-6, LM3) but not in normal hepatocytes AML12 (Fig. 4E).

Western blot (WB) analysis of sP10-treated SK-Hep1 cells revealed reduced NPM1 protein levels (Fig. 5A).

quantitative PCR showed no significant changes in NPM1 mRNA expression (Fig. 5B).

WB analysis demonstrated accelerated NPM1 degradation (shorter half-life) in sP10-treated cells (Fig. 5C).

if the cells were treated with the combination of CHX and MG132 or CHX and CQ, both could prevent the decrease in NPM1 protein level, and especially CQ significantly upregulated the level of NPM1 protein (Fig. 5D).

Western blot analysis showed that both sP10 and sP177 inhibited the expression of α-SMA and Collagen I in a dose-dependent manner (Fig. 2D).

Transwell assays showed that the migration ability of LX-2 cells was downregulated by intracellular expression of peptide aptamers (P10 and P177) and incubation of synthetic peptides (sP10 and sP177) (Fig. 2B).

The results of the Transwell assay (Fig. 3C) and wound healing experiments (Supplementary Fig. S4A) indicated that P10 and sP10 inhibited the migration and invasion of SK-Hep1 and Hepa1-6 cells, respectively.

The results of the Transwell assay (Fig. 3C) and wound healing experiments (Supplementary Fig. S4A) indicated that P10 and sP10 inhibited the migration and invasion of SK-Hep1 and Hepa1-6 cells, respectively.

the clonal formation assay showed that P10 and sP10 decreased the number of clones formed by SK-Hep1 and Hepa1-6 cells (Fig. 3B).

the clonal formation assay showed that P10 and sP10 decreased the number of clones formed by SK-Hep1 and Hepa1-6 cells (Fig. 3B).

sP10 unexpectedly reduced ROS in SK-Hep1 HCC cells (Fig. S7).

sP10/sP177 moderately increased ROS levels in LX-2 hepatic stellate cells (consistent with NPM1 downregulation) (Fig. S7).

The proliferation assay results indicated that when the expression of NPM was significantly downregulated, sP10 no longer inhibited cell proliferation (Supplementary Fig. S3B).