Journal: STAR Protocols

Article Title: Protocol for enhancing CRISPR-Cas9 genome editing using histone deacetylase inhibition and engineered virus-like particle delivery

doi: 10.1016/j.xpro.2026.104493

Figure Lengend Snippet: FAME-CRISPR improves CRISPR/Cas9 gene editing efficiency through HDACi mediated chromatin relaxation and precise eVLP delivery (A) shows a brief schematic of the major steps required to improve editing efficacy using FAME-CRISPR workflow. (B), (C), (D), and (E) show representative results obtained from MDA-MB-231 one of the utilized cell lines from the original FAME-CRISPR study ([60nM panobinostat] and [1.5 μM belinostat]) (n = 3 biological replicates for all conditions, and error bars reflect standard deviation). Results and raw data for MDA-MB-231 and other tested cell lines are available in the original study and data availability statement section of STAR Protocols.

Article Snippet: MDA-MB-231 , ATCC , #HTB-26.

Techniques: CRISPR, Standard Deviation

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Ptch1 is expressed in breast cancer cell lines. A. PTCH1 mRNA expression (log 2 transformed) in various luminal and HER2 breast cancer cell lines (dark green) and in TNBC cell lines (other colors). TNBC cell lines are depicted according to the “Lehmann TNBC subtype” nomenclature : basal-like 1 (yellow), basal-like 2 (pale green), immunomodulatory (brown), luminal androgen receptor (dark pink), mesenchymal (pale pink) and mesenchymal stem-like (pink). B. PTCH1 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from TNBC cell lines (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against PTCH1. PTCH1 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transformation Assay, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 drug efflux inhibitor PAH increases the sensitivity of TNBC cells to chemotherapy . Cell viability was measured after 24 h or 48 h treatment with increasing concentration of docetaxel or doxorubicin respectively on MDA-MB-231, MDA-MB-468 and HCC-38 cell lines in the absence or the presence of 15µM PAH. IC 50 values (corresponding to the concentration of chemotherapy inducing 50% of cell death) were calculated. Data reported are the mean ± SEM of at least 3 independent experiments. Significance is attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Other multidrug transporters are expressed in TNBC cell lines. A. P-gp and ABCG2 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from each TNBC cell line (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against P-gp or ABCG2 and GAPDH. B. P-gp, ABCG2 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*). ** P < 0.01; *** P < 0.001; ns P > 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PAH increases docetaxel efficacy against TNBC spheroids. MDA-MB-231 cells were plated in ultra-low attachment surface 24 well plates in complete medium and treated with increasing concentrations of docetaxel in the presence of DMSO (control, 0 PAH), PAH 10 µM or 30 µM. After 2 weeks pictures were taken using Cytation 5 cell imaging system from Biotek. The surface of spheroids was calculated and reported after normalization on the condition without docetaxel for each concentration of PAH.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Control, Imaging, Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PAH addition to chemotherapy inhibits migration of TNBC cells. A. 100000 cells were seeded on membrane from Transwell plates. Cells were treated 24 h with doxorubicin ± 15µM PAH or 48 h with docetaxel ± 15µM PAH. After fixation and staining with crystal violet, wells were observed on a microscope with X5 objective. B. Migration was measured using a wound-healing assay. A wound was performed on MDA-MB-231 confluent cells seeded in 24 well plates. The medium was replaced by fresh medium containing 5 µM docetaxel in the absence of PAH, or the presence of 5 µM PAH. Two pictures were taken at two different points of each well immediately after wound, and 3 days after wound with 5X objective. The width of the wound was measured using ImageJ software and reported as final wound width/initial wound width in percentage. Data presented are the mean ± SEM of 3 independent experiments. Significance is attained at P < 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Migration, Membrane, Staining, Microscopy, Wound Healing Assay, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 contributes to the efflux of doxorubicin from TNBC cells. A. PTCH1 protein expression (left panel) and intracellular doxorubicin (right panel) were analyzed 16 h after transfection of MDA-MB-231 cells with PTCH1-siRNA or negative-control-siRNA. PTCH1 and GAPDH western blot signals were quantified using ImageJ software (left panel). For doxorubicin accumulation measurements (right panel), MDA-MB-231 cells were grown on slides and transfected with PTCH1-siRNA or negative-control-siRNA. After incubation with 10 µM doxorubicin, 3 coverslips were fixed for doxorubicin loading control; the other coverslips were incubated with efflux buffer and fixed. B. Docetaxel inhibits the accumulation of doxorubicin in MDA-MB-231 cells. Cells on coverslip were incubated with 10 µM doxorubicin or 10 µM doxorubicin and 50 µM docetaxel. Doxorubicin fluorescence images were acquired by epifluorescence microscopy using a 40X objective, and doxorubicin fluorescence was quantified using ImageJ software for about 100 cells per condition per experiment. Histograms are the mean ± SEM of 3 independent experiments. Significance is attained at P < 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transfection, Negative Control, Western Blot, Software, Incubation, Control, Fluorescence, Epifluorescence Microscopy

Journal: Materials Today Bio

Article Title: Mannosylated graphene oxide nanotherapeutics co-delivering docetaxel and a STING agonist reprogram myeloid cells and potentiate antitumor immunity

doi: 10.1016/j.mtbio.2026.103086

Figure Lengend Snippet: Macrophage-targeted GO-EDM-DTX-Vad induces M2-to-M1 repolarization and limits TNBC cell proliferation and migration. (A) After incubation of M2-polarized macrophages with the nanocomposites for 24 h, CD206 and CD86 were quantified by flow cytometry. (B) After co-treatment of M0 RAW264.7 cells with IL-4/IL-13 and the nanocomposites for 48 h, CD206 and CD86 were quantified by flow cytometry. (C) RT-qPCR of CD86, iNOS, CD206 and IL-10 expression. (D) RAW264.7 immunoblots of STING axis including TBK1, phospho-TBK1, IRF3 and phospho-IRF3, with GAPDH as a reference. (E) CLSM images of RAW264.7 uptake of RhoB–GO-EDM-DTX-Vad (nuclei, blue; nanocomposites, red; scale bar, 100 μm). (F) ELISA of TGF-β, IL-10 and TNF-α in culture supernatants. (G) Schematic of the in vitro workflow for generating macrophage-conditioned media and assessing CM-driven tumor cell functions. (H, I) Colony formation (H) and EdU flow-cytometry analyses (I) of 4T1 and MDA-MB-231 cells after treatment with the indicated macrophage-CM. Groups: G1, PBS; G2, GO-EDM; G3, GO-EDM-DTX; G4, GO-EDM-Vad; G5, GO-EDM-DTX-Vad; G6, GO-EDM-DTX-Vad + NIR. Data are mean ± SD (n = 3). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001; ns, not significant.

Article Snippet: 4T1 and MDA-MB-231 cells were treated with the indicated nanocomposites for 24 h. The apoptosis was quantified by Annexin V-FITC/PI staining (Elabscience, Wuhan, China) followed by flow cytometry, and cell viability was visualized by Calcein-AM/PI live/dead staining (Servicebio, Wuhan, China) using CLSM.

Techniques: Migration, Incubation, Flow Cytometry, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, In Vitro

Journal: Materials Today Bio

Article Title: Mannosylated graphene oxide nanotherapeutics co-delivering docetaxel and a STING agonist reprogram myeloid cells and potentiate antitumor immunity

doi: 10.1016/j.mtbio.2026.103086

Figure Lengend Snippet: GO-EDM-DTX-Vad combined with NIR induces TNBC cell death, apoptosis and immunogenic cell death. (A, B) Calcein-AM/PI live/dead staining of 4T1 (A) and MDA-MB-231 (B) cells (live, green; dead, red; scale bar, 200 μm). (C, D) Annexin V/PI apoptosis plots for 4T1 (C) and MDA-MB-231 (D) cells. (E, F) Quantification of viable and early/late apoptotic 4T1 cells. (H, I) Quantification of viable and early/late apoptotic MDA-MB-231 cells. (G, J) Dose–response viability curves after 48 h treatment with free DTX or GO-EDM-DTX-Vad in 4T1 (G) and MDA-MB-231 (J). (K, L) Immunofluorescence of CRT exposure and HMGB1 release after 24 h treatment in 4T1 (K) and MDA-MB-231 (L) cells (nuclei, blue; CRT/HMGB1, green; scale bar, 100 μm). (M) Schematic of the ICD-to-DC maturation assay. (N–P) Frequencies of CD40 + , CD80 + and CD86 + DCs after co-culture. Data are mean ± SD (n = 3). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001; ns, not significant.

Article Snippet: 4T1 and MDA-MB-231 cells were treated with the indicated nanocomposites for 24 h. The apoptosis was quantified by Annexin V-FITC/PI staining (Elabscience, Wuhan, China) followed by flow cytometry, and cell viability was visualized by Calcein-AM/PI live/dead staining (Servicebio, Wuhan, China) using CLSM.

Techniques: Staining, Immunofluorescence, Co-Culture Assay

Journal: Translational Oncology

Article Title: Toxicarioside H induces cytoprotective autophagy by hindering the progression of necroptosis in triple-negative breast cancer cells

doi: 10.1016/j.tranon.2026.102779

Figure Lengend Snippet: Effects of ToxH on TNBC cell proliferation and cell death. TNBC MDA-MB-231, MDA-MB-468, and HCC 38 cells were treated with different concentrations of ToxH as indicated for 24 h. (A) Cell viability was assessed using the CCK-8 assay, represented as a percentage relative to the control group (Ctrl). (B) Representative images from the EdU assay showing proliferating cells (red fluorescence). (C) Quantification of EdU-positive proliferating cells by flow cytometry, represented as a percentage relative to the control group (Ctrl). (D) Representative flow cytometry images showing the intensity of 7-AAD-stained cells. (E) Percentage of 7-AAD-positive dead cells determined by flow cytometry. (F) Measurement of LDH release, represented as fold change relative to the control group (Ctrl). Data are presented as mean ± SD and analyzed using one-way ANOVA with Tukey's multiple comparisons test, N = 5; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; ns, not significant.

Article Snippet: The TNBC cell lines used in this study, including MDA-MB-231, MDA-MB-436, and HCC 38, were obtained from the American Type Culture Collection (ATCC) and maintained in our laboratory.

Techniques: CCK-8 Assay, Control, EdU Assay, Fluorescence, Flow Cytometry, Staining

Journal: Translational Oncology

Article Title: Toxicarioside H induces cytoprotective autophagy by hindering the progression of necroptosis in triple-negative breast cancer cells

doi: 10.1016/j.tranon.2026.102779

Figure Lengend Snippet: ToxH triggers the complete autophagic process in TNBC cells. MDA-MB-231, MDA-MB-438, and HHC-38 cells were treated with or without ToxH for 24 h and transfected with or without the corresponding plasmid. (A) Western blotting detection shows an increase in the expression of LC3-II, ATG5, and Beclin-1, but a decrease in the expression of p62. (B) Immunofluorescence observation reveals that ToxH treatment generated a considerable number of LC3B puncta. (C) TNBC cells transfected with the mCherry-GFP-LC3 plasmid demonstrate an observable increase in the formation of autophagosomes (yellow puncta) and autolysosomes (red puncta) that were treated with ToxH. Data are presented as mean ± SD and analyzed using an Unpaired t-test (A) or two-way ANOVA with Tukey's multiple comparisons test (B and C), N = 3 (A) or 100 (B and C); * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; ns, not significant.

Article Snippet: The TNBC cell lines used in this study, including MDA-MB-231, MDA-MB-436, and HCC 38, were obtained from the American Type Culture Collection (ATCC) and maintained in our laboratory.

Techniques: Transfection, Plasmid Preparation, Western Blot, Expressing, Immunofluorescence, Generated

Journal: Translational Oncology

Article Title: Toxicarioside H induces cytoprotective autophagy by hindering the progression of necroptosis in triple-negative breast cancer cells

doi: 10.1016/j.tranon.2026.102779

Figure Lengend Snippet: Combination of ToxH and CQ treatments increases anti-tumor activity through promotion of necroptosis in TNBC cells. MDA-MB-231, MDA-MB-438, and HHC-38 tumor models were established in female nude mice and treated with ToxH or combined with CQ for five mice in each group. (A) Tumor images were captured at day 18 after tumor cell inoculation. (B) Tumor volumes were measured at day 18 after the tumor cell inoculation. (C) Western blotting detection of the autophagy marker proteins LC3 and p62, and necroptosis marker proteins RIPK1, RIPK3, and MLKL and phosphorylated MLKL (pMLKL) in the tumor tissues treated with the indicated reagents. (D) Immunohistochemical detection of the necroptosis marker phosphorylated proteins pRIPK3 and pMLKL. Data are presented as mean ± SD and were assessed using one-way ANOVA with Tukey's post hoc test, where N = 5 (A and B) or representative images from triplicate experiments (C and D); * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; ns, no significant.

Article Snippet: The TNBC cell lines used in this study, including MDA-MB-231, MDA-MB-436, and HCC 38, were obtained from the American Type Culture Collection (ATCC) and maintained in our laboratory.

Techniques: Activity Assay, Western Blot, Marker, Immunohistochemical staining

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Ptch1 is expressed in breast cancer cell lines. A. PTCH1 mRNA expression (log 2 transformed) in various luminal and HER2 breast cancer cell lines (dark green) and in TNBC cell lines (other colors). TNBC cell lines are depicted according to the “Lehmann TNBC subtype” nomenclature : basal-like 1 (yellow), basal-like 2 (pale green), immunomodulatory (brown), luminal androgen receptor (dark pink), mesenchymal (pale pink) and mesenchymal stem-like (pink). B. PTCH1 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from TNBC cell lines (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against PTCH1. PTCH1 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transformation Assay, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 drug efflux inhibitor PAH increases the sensitivity of TNBC cells to chemotherapy . Cell viability was measured after 24 h or 48 h treatment with increasing concentration of docetaxel or doxorubicin respectively on MDA-MB-231, MDA-MB-468 and HCC-38 cell lines in the absence or the presence of 15µM PAH. IC 50 values (corresponding to the concentration of chemotherapy inducing 50% of cell death) were calculated. Data reported are the mean ± SEM of at least 3 independent experiments. Significance is attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Other multidrug transporters are expressed in TNBC cell lines. A. P-gp and ABCG2 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from each TNBC cell line (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against P-gp or ABCG2 and GAPDH. B. P-gp, ABCG2 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*). ** P < 0.01; *** P < 0.001; ns P > 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Ptch1 is expressed in breast cancer cell lines. A. PTCH1 mRNA expression (log 2 transformed) in various luminal and HER2 breast cancer cell lines (dark green) and in TNBC cell lines (other colors). TNBC cell lines are depicted according to the “Lehmann TNBC subtype” nomenclature : basal-like 1 (yellow), basal-like 2 (pale green), immunomodulatory (brown), luminal androgen receptor (dark pink), mesenchymal (pale pink) and mesenchymal stem-like (pink). B. PTCH1 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from TNBC cell lines (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against PTCH1. PTCH1 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transformation Assay, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 drug efflux inhibitor PAH increases the sensitivity of TNBC cells to chemotherapy . Cell viability was measured after 24 h or 48 h treatment with increasing concentration of docetaxel or doxorubicin respectively on MDA-MB-231, MDA-MB-468 and HCC-38 cell lines in the absence or the presence of 15µM PAH. IC 50 values (corresponding to the concentration of chemotherapy inducing 50% of cell death) were calculated. Data reported are the mean ± SEM of at least 3 independent experiments. Significance is attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Other multidrug transporters are expressed in TNBC cell lines. A. P-gp and ABCG2 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from each TNBC cell line (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against P-gp or ABCG2 and GAPDH. B. P-gp, ABCG2 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*). ** P < 0.01; *** P < 0.001; ns P > 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Western Blot, Software