Journal: bioRxiv

Article Title: AP-1 regulates heterogeneous cellular dormancy in TNBC

doi: 10.1101/2023.11.22.566980

Figure Lengend Snippet: (a) SCCs were enriched in AP-1 subunits in mRNA level. Relative gene expressions of FACS sorted SCCs and bulk cells were analyzed by RT-qPCR. Data are represented as mean ± SD. (b) JunB, p21 and GADD45A were enriched in H2B-eGFP enriched SCCs after 12 days of chasing. Scale bar = 20 μm. (c) CC-930 could reduce SCCs residual caused by chemotherapy in vitro. TAC was a combination of docetaxel, doxorubicin, and cyclophosphamide, with common clinical dosage. Data are represented as mean ± SD. (d) mCherry were expressed via AP1RE to construct AP-1 transcriptional activity reporter system. AP-1 promoted mCherry and SCCs marker H2B-eGFP were co-localized after 12 days of chasing. Scale bar = 20 μm. (e) SCCs had higher AP-1 transcriptional activity, and Violet enriched SCCs’ was most robust. (f-g) CC-930 could reduce gene expression and protein level upregulated by TAC treatment. MDA-MB-231 cells were treated with TAC and 10 mM CC-930. Relative gene expressions were analyzed by RT-qPCR (f) , and protein level were analyzed by Western Blot (g) . Data are represented as mean ± SD. *p ≤0.05; **p ≤0.01; ***p ≤0.001, Student’s t test.

Article Snippet: A combination of 1.867 μM docetaxel, 1.724 μM doxorubicin, and 35.829 μM cyclophosphamide (TargetMol.USA) was named as TAC.

Techniques: Quantitative RT-PCR, In Vitro, Construct, Activity Assay, Marker, Expressing, Western Blot

Journal: Experimental and Therapeutic Medicine

Article Title: Circular RNA expression profiling and the potential role of hsa_circ_0005379 in decreased ovarian reserve

doi: 10.3892/etm.2025.12974

Figure Lengend Snippet: Function of hsa_circ_0005379 in CTX-induced KGN ovarian granulosa cells. (A) Different gradient concentrations (0, 10, 15, 20, 25 and 30 µM) of CTX were used to induce ovarian granulosa cell damage, and Cell Counting Kit-8 assay was used to detect KGN cell viability. (B) RT-qPCR analysis of hsa_circ_0005379 expression after transfection with three shRNAs and negative control sh-NC. (C) RT-qPCR analysis of hsa_circ_0005379 expression after transfection with OE vector and an empty OE-NC vector. (D) The apoptosis of sh-hsa_circ_0005379 or hsa_circ_0005379-OE transfected KGN cells was detected using annexin V/PI staining. (E) Flow cytometry was used to detect the ROS level of cells after different treatments. The concentration of (F) MDA and (G) SOD in KGN cells with different treatments was determined using corresponding biochemistry detection kits. n=3, * P<0.05, ** P<0.01 and *** P<0.001 vs. 0 µΜ or as indicated. CTX, cyclophosphamide; RT-qPCR, reverse transcription-quantitative PCR; OE, overexpression; NC, negative control; sh, short hairpin; SOD, superoxide dismutase; MDA, malondialdehyde; ROS, reactive oxygen species; MFI, mean fluorescence intensity; ns, not significant.

Article Snippet: Cyclophosphamide (CTX; cat. no. HY-17420; MedChemExpress) may destroy the follicular pool, leading to primary ovarian insufficiency ( ).

Techniques: Cell Counting, Quantitative RT-PCR, Expressing, Transfection, Negative Control, Plasmid Preparation, Staining, Flow Cytometry, Concentration Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Over Expression, Fluorescence

Journal: Cell Death Discovery

Article Title: The EZH2-H3K27me3 axis modulates aberrant transcription and apoptosis in cyclophosphamide-induced ovarian granulosa cell injury

doi: 10.1038/s41420-023-01705-6

Figure Lengend Snippet: A Western blot results of the H3K27me3, H2AK119ub1, H3K9me3, H3K4me3, EZH2, and SUZ12 levels in the PBS and CTX groups. H3, GAPDH, and β-tubulin were blotted as loading controls. N = 6 mice per time point for each group. B – D Immunofluorescence staining of the ovaries collected 6 and 24 h after the i.p. injection of PBS or CTX. Rabbit monoclonal antibodies H3K27me3 ( B ), H3K9me3 ( C ), and EZH2 ( D ) were detected using anti-rabbit IgG (red). TUNEL was probed with Alexa Fluor 488 (green). Cell nuclei were labeled with DAPI (blue). Scale bar, 100 μm. N = 6 ovaries from different mice per time point for each group. E Western blot results for EZH2, H3K27me3, and γH2AX levels in the negative control (NC) and 4-HC treatment groups. β-actin and H3 were blotted as loading controls. F Quantitative expression of Ezh2 in the siNC and si Ezh2 -1 + 2 + 3 groups. G Western blot results of EZH2, H3K27me3, and γH2AX levels in the siNC and si Ezh2 -1 + 2 + 3 groups. β-actin and H3 were blotted as loading controls. Data are the mean ± SD of at least three independent experiments. Statistical analyses were carried out using a two-tailed Student’s t -test; *** P < 0.001.

Article Snippet: To evaluate the direct role of CTX, the GCs were treated with 2.5 μM 4-HC (the in vitro active form of CTX, HY-117433; MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Western Blot, Immunofluorescence, Staining, Injection, Bioprocessing, TUNEL Assay, Labeling, Negative Control, Expressing, Two Tailed Test

Journal: Oncotarget

Article Title: Transplantation of autologous bone marrow pre-loaded ex vivo with oncolytic myxoma virus is efficacious against drug-resistant Vk*MYC mouse myeloma

doi: 10.18632/oncotarget.28205

Figure Lengend Snippet: ( A ) Diagram describing the experimental design including the treatment time-line after myeloma implantation. ( B ) Survival curves of different cohorts comparing the therapeutic effects of 100 mg/Kg cyclophosphamide (Cy) alone versus Cy followed by transplant with autologous BM pre-loaded with un-armed vMyxM135KO virus versus Cy followed by transplant with autologous BM pre-loaded with armed vMyx-hTNF virus. (B) Survival curves show the number of treated mice (n), survival (in days) from the beginning of cancer implantation, and P values for log-rank Mantel-Cox survival test. P values are reported as not statistically significant (ns) when P > 0.05 or statistically significant when **** P ≤ 0.0001.

Article Snippet: In brief, cyclophosphamide (Cy) (Tocris Bioscience) was used at a final concentration of 100 mg/Kg and administrated intraperitoneally (i.p.) twice, each dose one week apart.

Techniques: Virus