Fig 2: PDBAG1 promotes ubiquitin‐dependent degradation of C1QBP. (A) The protein level of C1QBP and HIF‐1α were assessed by immunoblotting in TNBC cells after treated with PDBAG1 within 12 h. (B) C1QBP protein level was assessed by immunoblotting after treated with PDBAG1 within 4 h. (C) The degradation speed of C1QBP protein was performed with protein extracted at 0, 4, 8, 16 and 24 h after treatment of MDA‐MB‐231 cells with cycloheximide (CHX) and PDBAG1. (D) Treating TNBC cells with inhibitors of different protein degradation associated pathways stacked with PDBAG1, the ubiquitin–proteasome pathway inhibitors MG‐132 and BZ could reverse the decrease in C1QBP protein level (long expose) caused by PDBAG1 (CQ, chloroquine; LEU, leupeptin hemisulphate; Baf, bafilomycin A1; BZ, bortezomib). (E) 24 h after MDA‐MB‐231 cells were treated with PDBAG1 and MG132, the intracellular fluorescence intensity and distribution of C1QBP and mitotracker (thermos fisher) were detected by immunofluorescence. Quantification from three independent experiments is presented as histograms. Magnification: 40×, Scale bar, 20 µm (Leica STELLARIS STED). (F) The impact of PDBAG1 treatment on C1QBP ubiquitination was assessed over a 12‐h period using co‐immunoprecipitation (co‐IP) assays. Results demonstrate a significant enhancement of C1QBP ubiquitination upon exposure to PDBAG1.

Fig 3: Honey disrupts RNA Pol I subunits and c-Myc to modulate ribosome biogenesis in MIA PaCa-2 and AsPC-1 cells. The images were acquired after 24 h of honey exposure through Leica STELLARIS STED & STELLARIS 8 STED Microscope. Magnification scale bar represents 10 µm.

Fig 4: HSV-1 infected human corneal epithelial cells. (A) Progression of HSV-1 infection over 48 h, with virus visualized after staining with an anti-HSV-1 antibody (red) and DAPI (blue). Scale bars, 33 μm. (B) Infected HCECs stained with antibodies against F-actin (green), HSV-1 (red), TSG101 (yellow) and DAPI (blue), imaged on a Leica Stellaris STED super resolution microscope. Arrows indicate TNTs. Scale bars, 23 μm.

Fig 5: PDBAG1 inhibits TNBC by direct target and decrease C1QBP at protein level. (A) Schematic diagram of the biotin‐labelled PDBAG1 pull down assay. (B and C) The silver staining gel picture and mass spectrometry results of the pulldown assay. (D) Pulldown experiment to verify the top five high abundant proteins bound by PDBAG1. (E and F) Schematic diagram and the results of the co‐IP assay (HA‐tagged C1QBP were used as bait, and biotin‐labelled PDBAG1 was added to the protein lysate). (G and H) Protein level of C1QBP was tested in TNBC cell lines after treated with PDBAG1 and change of C1QBP in the cytoplasm and nucleus of MDA‐MB‐231 cells after treated with PDBAG1. (I) 24 h after MDA‐MB‐231 cells were treated with PDBAG1, the intracellular fluorescence intensity and distribution of C1QBP and mitotracker (Thermo Fisher) were detected by immunofluorescence. Quantification of C1QBP from three independent experiments is presented as histograms. Magnification: 40×, Scale bar, 20 µm (Leica STELLARIS STED). (J) Stable overexpression C1QBP MDA‐MB‐231 cell lines were constructed to detect C1QBP protein level after treated with PDBAG1. (K, L and M) Stable overexpression tumour cells were inoculated into nude mice for tumourigenic model experiments (n = 6 mice in each group) were constructed, after treated with vehicle or PDBAG1 the effects were shown on tumour image, tumour volume and tumour weight were examined (NS, non‐significant, *p < .05, **p < .01, ***p < .001; two‐way ANOVA test. Error bars, mean ± SD, n = 6). (N and O) The expression levels of C1QBP in different subtypes of breast cancer in the TCGA database and the effect of high expression of C1QBP on DFS of patients with all types of breast cancer and triple negative breast cancer (GEPIA2, http://gepia2.cancer‐pku.cn).