Journal: Materials Today Bio

Article Title: Toll-like receptor 9-overexpressing red blood cell biomimetic microbubbles adsorb cell-free DNA to relieve steatotic liver ischemia-reperfusion injury

doi: 10.1016/j.mtbio.2026.103558

Figure Lengend Snippet: Synthetic RBC-MBs with high TLR9 expression (A-B) TLR9 protein levels on erythrocyte membranes were significantly decreased in both patients with MASLD (A), n = 9/7, and MASLD rats (B), n = 8/6. Erythrocytes were collected from patients with or without MASLD (A) and from rats fed a normal diet or a high-fat diet (B). Erythrocyte membranes were isolated and subjected to western blotting ( upper panels ). TLR9 levels were normalized to ATP1A levels in each sample, and the normalized values were used for statistical analysis ( lower panels ). (C) Lipopolysaccharide (LPS) treatment significantly increased TLR9 protein levels on erythrocyte membranes. Rats were intravenously injected with 12 mg kg -1 LPS for 6 h. Erythrocytes were collected, and the membranes were isolated and analyzed by western blotting ( upper panel ). TLR9 levels were normalized to ATP1A levels in each sample, and the normalized values were used for statistical analysis ( lower panel ), n = 6. (D) Schematic diagram illustrating the synthesis of RBC-MBs. (E) The ultrasound microbubbles appeared as a milky suspension. (F-G) Size distribution of Con-MBs (F) and RBC-MBs (G) in phosphate-buffered saline. (H-I) Average hydrodynamic diameters (H) and surface charges (ζ-potential) (I) of Con-MBs and RBC-MBs, as determined by dynamic light scattering, n = 6. (J) RBC-MBs were round, uniform, and well-dispersed. Erythrocyte membranes were labeled with DiL (red), and phospholipids were labeled with DiO (green). Scale bar: 30 μm. (K) RBC-MBs contained TLR9, CD47, and ATP1A proteins. Red blood cells, erythrocyte membranes, and RBC-MBs were subjected to western blotting. (L) RBC-MBs effectively adsorbed cfDNA in vitro . BRL-3A cells were treated with 250 μM sodium palmitate and 500 μM sodium oleate for 24 h, followed by hypoxia for 9 h and reoxygenation for 2 h. Subsequently, the cells were co-incubated with DiO-labeled (green) Con-MBs or RBC-MBs for 20 min, then immunostained with an anti-DNA antibody (red) and DAPI (blue, nuclei), and imaged by fluorescence microscopy. Scale bar: 20 μm. For (A-C, H-I), error bars represent mean ± standard deviation. P values were calculated using the unpaired Student's t-test (A, B, C, H, I). *, P < 0.05; ***, P < 0.001; ****, P < 0.0001.(For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary antibodies used included: rabbit polyclonal antibodies against TLR9 (A14642, Abclonal), NF-κB (A2547, Abclonal), MyD88 (A0980, Abclonal), cGAS (A8335, Abclonal), IL-1α (A2170, Abclonal), TNF-α (A0277, Abclonal), IL-6 (A0286, Abclonal), CD206 (A8301, Abclonal), CD86 (A16805, Abclonal), CD47 (A1838, Abclonal), and ATP1A (A0643, Abclonal); rabbit monoclonal antibodies against STING (13647, CST, Beverly, MA, USA), p-NF-κB (3033, CST), p-TBK1 (5483, CST), TBK1 (ab40676, Abcam), p-IκB (2859, CST), IκB (4812, CST), p-IRF3 (4947, CST), and β-actin (AC026, Abclonal).

Techniques: Expressing, Isolation, Western Blot, Injection, Suspension, Saline, Labeling, In Vitro, Incubation, Fluorescence, Microscopy, Standard Deviation

Journal: Materials Today Bio

Article Title: Toll-like receptor 9-overexpressing red blood cell biomimetic microbubbles adsorb cell-free DNA to relieve steatotic liver ischemia-reperfusion injury

doi: 10.1016/j.mtbio.2026.103558

Figure Lengend Snippet: cfDNA promotes the polarization of macrophages towards the M1 phenotype by activating the TLR9-MyD88 and cGAS-STING pathways (A- D) CpG DNA treatment increased the protein levels of IL-1α, TNF-α, and IL-6. RAW264.7 cells were treated with 1 μg mL -1 CpG DNA for 24 h and then subjected to western blotting (A). IL-1α (B), TNF-α (C), and IL-6 (D) levels were normalized to the β-actin level in each sample, and the normalized values were used for statistical analysis, n = 4. (E-F) CpG DNA treatment significantly enhanced CD86 expression in macrophages. RAW264.7 cells were treated with 1 μg mL -1 CpG DNA for 24 h, then immunostained with an anti-CD86 antibody (green) and DAPI (blue, nuclei), and imaged by fluorescence microscopy. Representative images (E) and quantitative fluorescence graph (F) are shown, n = 6. Scale bar: 50 μm. (G-J) CpG DNA treatment significantly upregulated the expression of key proteins associated with the TLR9-MyD88 and cGAS-STING pathways. RAW264.7 cells were treated with 1 μg mL -1 CpG DNA for 24 h and then subjected to western blotting, n = 8/4(G, I). STING, cGAS, TLR9, and MyD88 levels were normalized to the β-actin level in each sample. p-NF-κB, p-IκB, p-TBK1, and p-IRF3 levels were normalized to their respective total protein levels (NF-κB, IκB, TBK1, and IRF3) in each sample. These normalized values were then used for statistical analysis, n = 8/4(H, J). For (B-D, F, H, J), error bars represent mean ± standard deviation. P values were calculated using one-way analysis of variance (ANOVA) (B-D, F, H, J). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.(For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary antibodies used included: rabbit polyclonal antibodies against TLR9 (A14642, Abclonal), NF-κB (A2547, Abclonal), MyD88 (A0980, Abclonal), cGAS (A8335, Abclonal), IL-1α (A2170, Abclonal), TNF-α (A0277, Abclonal), IL-6 (A0286, Abclonal), CD206 (A8301, Abclonal), CD86 (A16805, Abclonal), CD47 (A1838, Abclonal), and ATP1A (A0643, Abclonal); rabbit monoclonal antibodies against STING (13647, CST, Beverly, MA, USA), p-NF-κB (3033, CST), p-TBK1 (5483, CST), TBK1 (ab40676, Abcam), p-IκB (2859, CST), IκB (4812, CST), p-IRF3 (4947, CST), and β-actin (AC026, Abclonal).

Techniques: Western Blot, Expressing, Fluorescence, Microscopy, Standard Deviation

Journal: Materials Today Bio

Article Title: Toll-like receptor 9-overexpressing red blood cell biomimetic microbubbles adsorb cell-free DNA to relieve steatotic liver ischemia-reperfusion injury

doi: 10.1016/j.mtbio.2026.103558

Figure Lengend Snippet: RBC-MBs promotes the polarization of macrophages towards the M2 phenotype by inhibiting the TLR9-MyD88 and cGAS-STING pathways (A-F) RBC-MB treatment significantly attenuated CpG DNA-induced upregulation of pro-inflammatory cytokines (A-B), key proteins in the TLR9-MyD88 (C-D) and cGAS-STING (E-F) pathways. RAW264.7 cells were treated with 1 μg mL -1 CpG DNA for 24 h, followed by incubation with 0.5 mL RBC-MBs (concentration: 1 × 10 8 particles mL -1 ) for 24 h, and then subjected to western blot analysis (A, C, E). IL-1α, TNF-α, IL-6, TLR9, MyD88, STING, and cGAS levels were normalized to the β-actin level in each sample. p-NF-κB, p-TBK1, and p-IRF3 levels were normalized to their respective total protein levels (NF-κB, TBK1, and IRF3) in each sample. These normalized values were then used for statistical analysis (B, D, F), n = 6. (G) RBC-MB treatment significantly attenuated CpG DNA-induced upregulation of CD86 in macrophages. RAW264.7 cells were treated with 1 μg mL -1 CpG DNA for 24 h, followed by incubation with 0.5 mL RBC-MBs (concentration: 1 × 10 8 particles mL -1 ) for 24 h, then immunostained with an anti-CD86 antibody (green) and DAPI (blue, nuclei), and imaged by fluorescence microscopy. Representative images (left panel) and quantitative fluorescence graph (right panel) are shown. Scale bar: 50 μm, n = 6. (H-J) RBC-MB treatment significantly inhibited hepatic IRI-induced polarization of macrophages toward the M1 phenotype (H) and activation of the TLR9-MyD88 and cGAS-STING pathways (I-J). MASLD rats underwent 45 min of hepatic ischemia followed by reperfusion. At 0, 6, 12, 18, and 24 h post-reperfusion, the rats were treated with either Con-MBs or RBC-MBs. Primary macrophages were isolated from rat livers, then co-stained with CD68 and CD163 followed by flow cytometric analysis (H), n = 3/4, or subjected to western blotting (I-J). CD206, iNOS, TLR9, MyD88, STING, and cGAS levels were normalized to the β-actin level in each sample, and these normalized values were used for statistical analysis (J), n = 3. For (B, D, F, G right panel , H right panel , J), error bars represent mean ± standard deviation. P values were calculated using the unpaired Student's t-test (B, D, F, G right panel ) or one-way analysis of variance (ANOVA) (H right panel , J). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.(For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary antibodies used included: rabbit polyclonal antibodies against TLR9 (A14642, Abclonal), NF-κB (A2547, Abclonal), MyD88 (A0980, Abclonal), cGAS (A8335, Abclonal), IL-1α (A2170, Abclonal), TNF-α (A0277, Abclonal), IL-6 (A0286, Abclonal), CD206 (A8301, Abclonal), CD86 (A16805, Abclonal), CD47 (A1838, Abclonal), and ATP1A (A0643, Abclonal); rabbit monoclonal antibodies against STING (13647, CST, Beverly, MA, USA), p-NF-κB (3033, CST), p-TBK1 (5483, CST), TBK1 (ab40676, Abcam), p-IκB (2859, CST), IκB (4812, CST), p-IRF3 (4947, CST), and β-actin (AC026, Abclonal).

Techniques: Incubation, Concentration Assay, Western Blot, Fluorescence, Microscopy, Activation Assay, Isolation, Staining, Standard Deviation

Journal: Cellular Oncology

Article Title: NAA50-mediated SLU7 stabilization promotes cisplatin resistance in bladder cancer via regulating MAP3K3 mRNA nuclear export and p38 MAPK activation

doi: 10.1007/s13402-026-01219-0

Figure Lengend Snippet: SLU7 knockdown enhances cisplatin sensitivity in cisplatin-resistant BLCA cells. ( A ) Kaplan-Meier analysis of cancer-specific survival (CSS) in BLCA patients from the GSE169455 cohort stratified by SLU7 expression. ( B ) Kaplan-Meier analysis of recurrence-free survival (RFS) in BLCA patients from the GSE169455 cohort stratified by SLU7 expression. ( C ) Kaplan-Meier analysis of RFS in BLCA patients from the GSE87304 cohort stratified by SLU7 expression. ( D ) Western blotting analysis of SLU7 protein expression in parental BLCA cells (T24-WT, UMUC3-WT) and their cisplatin-resistant derivatives (T24-CIS, UMUC3-CIS). ( E ) Western blotting validation of SLU7 knockdown in T24-CIS and UMUC3-CIS cells. ( F ) Western blotting validation of SLU7 overexpression in T24 and UMUC3 cells. ( G - J ) CCK-8 assays showing cisplatin IC₅₀ values in T24-CIS and UMUC3-CIS cells transfected with shSLU7#2/4 ( G - H ) and SLU7-Flag-transfected T24 and UMUC3 cells ( I - J ). Data are presented as mean ± SD. p -values were calculated using unpaired two-tailed Student’s t-test or one-way ANOVA. *p < 0.05, ***p < 0.001, ****p < 0.0001

Article Snippet: The site-specific polyclonal antibody against SLU7 acetylated at serine 2 (SLU7^S2ac) was generated by ABclonal using a synthetic acetylated serine-containing peptide as the immunogen.

Techniques: Knockdown, Expressing, Western Blot, Biomarker Discovery, Over Expression, CCK-8 Assay, Transfection, Two Tailed Test

Journal: Cellular Oncology

Article Title: NAA50-mediated SLU7 stabilization promotes cisplatin resistance in bladder cancer via regulating MAP3K3 mRNA nuclear export and p38 MAPK activation

doi: 10.1007/s13402-026-01219-0

Figure Lengend Snippet: SLU7 promotes cisplatin resistance by activating the p38 MAPK pathway. ( A ) GO pathway enrichment analysis of RNA-seq data from SLU7-knockdown T24-CIS cells, highlighting downregulated MAP3K family signaling pathways. ( B ) Western blotting analysis of phosphorylation levels of ERK1/2, p38 MAPK, JNK/SAPK, and ERK5 in T24-CIS, UMUC3-CIS, and UMUC3 cells with SLU7 knockdown or overexpression. (C, E) Western blotting analysis of p38 and p-p38 protein levels in SLU7-overexpressing T24 ( C ) and UMUC3 ( E ) cells treated with the p38 inhibitor Adezmapimod. ( D , F ) CCK-8 assays showing cisplatin IC₅₀ values in SLU7-overexpressing T24 ( D ) and UMUC3 ( F ) cells treated with Adezmapimod. Data are presented as mean ± SD. p -values were calculated using one-way ANOVA. *** p < 0.001, **** p < 0.0001, ns indicates no significant difference

Article Snippet: The site-specific polyclonal antibody against SLU7 acetylated at serine 2 (SLU7^S2ac) was generated by ABclonal using a synthetic acetylated serine-containing peptide as the immunogen.

Techniques: RNA Sequencing, Knockdown, Protein-Protein interactions, Western Blot, Phospho-proteomics, Over Expression, CCK-8 Assay

Journal: Cellular Oncology

Article Title: NAA50-mediated SLU7 stabilization promotes cisplatin resistance in bladder cancer via regulating MAP3K3 mRNA nuclear export and p38 MAPK activation

doi: 10.1007/s13402-026-01219-0

Figure Lengend Snippet: SLU7 upregulates MAP3K3 to activate the p38 MAPK pathway. ( A ) Western blotting analysis of MAP3K3 protein expression in T24-CIS, UMUC3-CIS, and UMUC3 cells with SLU7 knockdown or overexpression. ( B – D ) qRT-PCR analysis of MAP3K3 mRNA levels in T24-CIS ( B ), UMUC3-CIS ( C ), and UMUC3 ( D ) cells with SLU7 knockdown or overexpression. ( E , G ) Western blotting analysis of SLU7, MAP3K3, p38, and p-p38 protein levels in SLU7-overexpressing T24 ( E ) and UMUC3 ( G ) cells with MAP3K3 knockdown. ( F , H ) CCK-8 assays showing cisplatin IC₅₀ values in SLU7-overexpressing T24 ( F ) and UMUC3 ( H ) cells with MAP3K3 knockdown. Data are presented as mean ± SD. p -values were calculated using unpaired two-tailed Student’s t-test or one-way ANOVA. **p < 0.01, ***p < 0.001, ****p < 0.0001 , ns indicates no significant difference

Article Snippet: The site-specific polyclonal antibody against SLU7 acetylated at serine 2 (SLU7^S2ac) was generated by ABclonal using a synthetic acetylated serine-containing peptide as the immunogen.

Techniques: Western Blot, Expressing, Knockdown, Over Expression, Quantitative RT-PCR, CCK-8 Assay, Two Tailed Test

Journal: Cellular Oncology

Article Title: NAA50-mediated SLU7 stabilization promotes cisplatin resistance in bladder cancer via regulating MAP3K3 mRNA nuclear export and p38 MAPK activation

doi: 10.1007/s13402-026-01219-0

Figure Lengend Snippet: SLU7 mediates MAP3K3 mRNA nuclear export to regulate its expression. ( A ) RIP-qPCR analysis showing the interaction between SLU7 and MAP3K3 mRNA in UMUC3–SLU7-Flag cells. ( B , C ) The location of SLU7 and MAP3K3 mRNA in UMUC3 cells were analyzed using IF and smiFISH. Scale bar: 10 μm. The fluorescence intensity curves of DAPI (blue), SLU7 (green) and MAP3K3 mRNA (purple) that crossed the arrows were analyzed. ( D , E ) qRT-PCR analysis of MAP3K3 mRNA levels in nuclear and cytoplasmic fractions of T24-CIS ( D ) and UMUC3-CIS ( E ) cells with SLU7 knockdown. ( F , G ) RNA-FISH images showing subcellular localization of MAP3K3 mRNA (purple) in T24-CIS ( F ) and UMUC3-CIS ( G ) cells with SLU7 knockdown. Scale bar: 10 μm. Data are presented as mean ± SD. p -values were calculated using unpaired two-tailed Student’s t-test or one-way ANOVA. ****p < 0.0001 , ns indicates no significant difference

Article Snippet: The site-specific polyclonal antibody against SLU7 acetylated at serine 2 (SLU7^S2ac) was generated by ABclonal using a synthetic acetylated serine-containing peptide as the immunogen.

Techniques: Expressing, Fluorescence, Quantitative RT-PCR, Knockdown, Two Tailed Test

Journal: Cellular Oncology

Article Title: NAA50-mediated SLU7 stabilization promotes cisplatin resistance in bladder cancer via regulating MAP3K3 mRNA nuclear export and p38 MAPK activation

doi: 10.1007/s13402-026-01219-0

Figure Lengend Snippet: NAA50 maintains SLU7 protein stability via N-terminal acetylation. ( A ) MS analysis identifying N-terminal acetylation of SLU7. ( B ) Western blotting analysis of SLU7 protein expression in T24-CIS cells with knockdown of different NAT subtypes. ( C ) Western blotting analysis of SLU7, MAP3K3, p38, and p-p38 protein levels in T24-CIS, UMUC3-CIS, and UMUC3 cells with NAA50 knockdown or overexpression. ( D ) Western blotting analysis of SLU7, MAP3K3, p38, and p-p38 protein levels in T24-CIS and UMUC3-CIS cells treated with the specific NAA50 inhibitor Naa50-IN-1. ( E ) Western blotting analysis verifying that SLU7 knockdown abrogates the regulation effect of NAA50 on MAP3K3 and p38 phosphorylation in T24-CIS cells. ( F ) Cycloheximide chase assay detecting the time-dependent of SLU7 protein stability in UMUC3 cells with NAA50 knockdown (siNAA50) or NAA50 inhibitor (Naa50-IN-1) treatment, compared with their respective control groups(siNC or DMSO). (G and H) IP assays detecting SLU7^S2ac (N-terminal acetylation) levels in UMUC3 cells with NAA50 knockdown ( G ) or Naa50-IN-1 treatment ( H ) in the presence of the proteasome inhibitor MG132 pretreatment. ( I ) Western blotting analysis of SLU7 protein stability in UMUC3 cells expressing Flag-tagged SLU7 wild-type (SLU7 WT -Flag) or acetylation site-mutant (SLU7 MUT -Flag) constructs upon Naa50-IN-1 treatment with CHX pretreatment. ( J ) Ubiquitination assay in 293T cells overexpressing Flag-tagged SLU7, detecting SLU7 ubiquitination level upon Naa50-IN-1 treatment

Article Snippet: The site-specific polyclonal antibody against SLU7 acetylated at serine 2 (SLU7^S2ac) was generated by ABclonal using a synthetic acetylated serine-containing peptide as the immunogen.

Techniques: Western Blot, Expressing, Knockdown, Over Expression, Phospho-proteomics, Control, Mutagenesis, Construct, Ubiquitin Proteomics

Journal: Cellular Oncology

Article Title: NAA50-mediated SLU7 stabilization promotes cisplatin resistance in bladder cancer via regulating MAP3K3 mRNA nuclear export and p38 MAPK activation

doi: 10.1007/s13402-026-01219-0

Figure Lengend Snippet: Schematic diagram of the “NAA50–SLU7–MAP3K3-p38 MAPK” regulatory axis in cisplatin resistance. NAA50 catalyzes N-terminal acetylation of SLU7 to maintain its protein stability. Stabilized SLU7 directly binds to MAP3K3 mRNA, promoting its nuclear export and translation. Increased MAP3K3 protein activates the p38 MAPK pathway, leading to cisplatin resistance in BLCA. Targeting NAA50 with Naa50-IN-1 downregulates SLU7 expression, inhibits the MAP3K3-p38 MAPK pathway, and restores cisplatin sensitivity

Article Snippet: The site-specific polyclonal antibody against SLU7 acetylated at serine 2 (SLU7^S2ac) was generated by ABclonal using a synthetic acetylated serine-containing peptide as the immunogen.

Techniques: Expressing