Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A Venn diagram illustrating the screening logic for SLC7A5. Integrated analysis of GSE153867 , GSE26712 , and OS databases identified nine genes highly expressed in drug-resistant cells with intersecting prognostic values. B Gene Expression Analysis. Analysis of the GSE153867 dataset revealed elevated SLC7A5 expression in Olaparib-resistant A2780 cells (C1–C8) compared to their parental controls (O1–O8). C WB. Western blot revealed higher SLC7A5 expression in HEY and SK-OV-3 cells compared to IOSE80, and lower expression in CAOV-3 and OVCAR-8 cells. D , E Drug Resistance Assay. IC50 values showed resistance to olaparib with high SLC7A5 expression. F WB. Western blot showed significant increases in SLC7A5 expression in olaparib-resistant cells. G Colony-formation assay. Knockdown of SLC7A5 significantly reduced Olaparib resistance in resistant cells. Left. HEY-R. Right. SK-OV-3-R. Plating 1000 cells per well with drug concentrations set at their respective IC50 values. H Statistical graph of Edu Assay. Edu incorporation assay showed that SLC7A5 knockdown significantly inhibited the proliferation of resistant cells treated with Olaparib. Edu incubation time was 2 h. Blue. DAPI, Green. Edu. Statistical graph displays Edu-positive rate in all cells. I Apoptosis Assay. SLC7A5 knockdown increased sensitivity to Olaparib in resistant cells. Cell apoptosis levels were detected using Annexin V and PI. J Colony-formation assay. The addition of leucine does not affect the regulation of olaparib resistance by SLC7A5. Plating 1000 cells per well with drug concentrations set at their respective IC50 values. Leucine (1 μM) treatment for 48 h. K Apoptosis Assay. SLC7A5’s regulation of olaparib resistance is leucine-independent. Cell apoptosis levels were detected using Annexin V and PI. L Statistical graph of Edu Assay. The addition of rapamycin does not affect the regulation of olaparib resistance by SLC7A5. Rapamycin (18.74 μM) treatment for 24 h. Edu incubation time was 2 h. Blue. DAPI, Green. Edu. Statistical graph displays Edu-positive rate in all cells. M Kaplan−Meier survival curves of nude mice implanted orthotopically with SK-OV-3 R cells. In a xenograft survival study conducted in nude mice, 5 million SK-OV-3 R cells were subcutaneously injected. Mice were divided into four groups, NC + vehivel, SLC7A5 + vehivel, NC + lue and SLC7A5 + leu. Once the tumors reached a volume of 50 mm³, each group was randomly divided into control and treatment subgroups. The treatment subgroup received olaparib every other day for a total of eight doses at 25 mg/kg. In accordance with animal ethics, mice were euthanized when tumor volumes reached 1000 mm³, which was recorded as the endpoint for survival. The median survival times were as follows. 40 days for the NC + vehivel group, 42.5 days for the NC + leu group, 20 days for the SLC7A5 + vehivel group, 17.5 days for the SLC7A5 + lue group (n = 10). N In vivo experiments. In vivo experiments demonstrated that SLC7A5 knockdown enhanced the inhibitory effect of Olaparib on ovarian cancer (n = 6). Upper panel. Schematic of in vivo treatment protocol. Lower panel. Tumor images. O Tumor weight (g). All in vitro findings are derived from a minimum of three independent experiments. Error bars denote the standard deviation. Statistical significance is represented as follows. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant in comparison to normal or control treatment.

Article Snippet: Prior to the staining process, the sections were deparaffinized and rehydrated, followed by heat-induced epitope retrieval using citrate buffer at 121 °C for 30 min. To inhibit endogenous peroxidase activity, sections were treated with 0.3% hydrogen peroxide in methanol for 30 min. Non-specific binding was mitigated by blocking with 10% normal bovine serum, after which the sections were incubated overnight at 4 °C with rabbit polyclonal antibodies targeting SLC7A5( 28670-1-AP, Proteintech ), ERBB2( 84046-1-RR, Proteintech ), and Ki67 ( 27309-1-AP, Proteintech ).

Techniques: Gene Expression, Expressing, Western Blot, Colony Assay, Knockdown, EdU Assay, Incubation, Apoptosis Assay, Injection, Control, In Vivo, In Vitro, Derivative Assay, Standard Deviation, Comparison

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A Immunohistochemistry (IHC). IHC revealed elevated SLC7A5 expression in late-stage patients, which was positively correlated with pathological grade. B Statistical graph of IHC. The correlation between SLC7A5 expression and tumor staging was analyzed in tissue microarray samples. Results showed that SLC7A5 expression increased with tumor progression. Stage I (3/8), Stage II (9/34), Stage III (36/64), and Stage IV (25/26). This suggests a potential role of SLC7A5 in advanced tumor development. C Survival Analysis. Kaplan-Meier analysis indicated that patients with high SLC7A5 expression had poorer prognoses and shorter survival times. D , E WB. The high expression efficiency of SLC7A5 and the knockdown efficiency using lentiviral shSLC7A5 #1 and #2 were detected in HEY-R and SK-OV-3-R cells. Overexpression was achieved using SLC7A5 plasmid in CAOV-3 and OVCAR-8 cells. F , G Colony-formation assay. SLC7A5 knockdown significantly inhibited cell proliferation, whereas overexpression promoted it. F HEY-R and SK-OV-3-R, G Caov-3 and Ovcar-8. Plating 2000 cells per well. H , I Apoptosis Assays. SLC7A5 knockdown increased apoptosis in cells, whereas overexpression SLC7A5 inhibited it. H HEY-R and SK-OV-3-R, I Caov-3 and Ovcar-8. J – L In vivo experiments. Subcutaneous tumor growth in nude mice with SK-OV-3-R cells showed that SLC7A5 knockdown inhibited tumor growth. J Bioluminescence Imaging. K Tumor volume (mm 3 ). M IHC Analysis. IHC showed a significant decrease in Ki67 expression following SLC7A5 knockdown (n = 6). All the in vitro data were obtained from at least three independent experiments. Error bars represent the standard deviation. Statistical significance is denoted as follows. * P < 0.05, ** P < 0.001, *** P < 0.0001, ns non-significant compared to the normal or control treatments.

Article Snippet: Prior to the staining process, the sections were deparaffinized and rehydrated, followed by heat-induced epitope retrieval using citrate buffer at 121 °C for 30 min. To inhibit endogenous peroxidase activity, sections were treated with 0.3% hydrogen peroxide in methanol for 30 min. Non-specific binding was mitigated by blocking with 10% normal bovine serum, after which the sections were incubated overnight at 4 °C with rabbit polyclonal antibodies targeting SLC7A5( 28670-1-AP, Proteintech ), ERBB2( 84046-1-RR, Proteintech ), and Ki67 ( 27309-1-AP, Proteintech ).

Techniques: Immunohistochemistry, Expressing, Microarray, Knockdown, Over Expression, Plasmid Preparation, Colony Assay, In Vivo, Imaging, In Vitro, Standard Deviation, Control

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A GSEA . The enrichment analysis results of gene SLC7A5 in the fatty acyl-CoA biosynthesis pathway highlight its potential significance. With a normalized enrichment score (NES) of 1.5658, a P-value of 0.031, and a false discovery rate (FDR) of 0.0927, it suggests that SLC7A5 may play a crucial role in this biosynthetic process, warranting further investigation and study. B Correlation Analysis. Analysis of TCGA database revealed a positive correlation between SLC7A5 and de novo lipid metabolism pathway-associated proteins ACLY, ACACA, and FASN, suggesting that SLC7A5 is involved in lipid metabolism remodeling. C ELISA. Measurements of Tri-Glycerides, Free Fatty Acids, and Total Cholesterol showed that SLC7A5 knockdown significantly inhibited lipid metabolism in HEY and SK-OV-3 cells. Cells (5 × 10 6 , lysed by sonication. D Lipid Droplet Staining. LD540 staining revealed reduced lipid droplets upon SLC7A5 knockdown. Red fluorescence was observed using a fluorescence microscope at an excitation wavelength of 537 nm. E Immunohistochemistry (IHC). IHC analysis of tumor samples from mice showing reduced ACLY expression following SLC7A5 knockdown. F ELISA. ELISA analysis revealed that SB 204990, an ACLY inhibitor, significantly inhibited lipid metabolism remodeling caused by SLC7A5 overexpression in CAOV-3 and OVCAR-8 cells. Cells were treated with 100 µM SB 204990 for 24 h. G Lipid Droplet Staining. LD540 staining shows a significant reduction in lipid droplet accumulation induced by SLC7A5 overexpression following treatment with SB 204990. H Western Blot Analysis. SB 204990 inhibited the upregulation of ACLY, ACACA, and FASN proteins induced by SLC7A5 overexpression. I – K SB 204990 treatment markedly suppressed the increased Olaparib resistance in CAOV-3 and OVCAR-8 cells caused by SLC7A5 overexpression. I Colony formation assay. J Cell viability assay. K Apoptosis Assay. All in vitro data were derived from at least three independent experiments. Error bars represent standard deviation. Statistical significance is indicated by the following symbols. * P < 0.05, ** P < 0.001, *** P < 0.0001, ns non-significant compared to the normal or control treatments.

Article Snippet: Prior to the staining process, the sections were deparaffinized and rehydrated, followed by heat-induced epitope retrieval using citrate buffer at 121 °C for 30 min. To inhibit endogenous peroxidase activity, sections were treated with 0.3% hydrogen peroxide in methanol for 30 min. Non-specific binding was mitigated by blocking with 10% normal bovine serum, after which the sections were incubated overnight at 4 °C with rabbit polyclonal antibodies targeting SLC7A5( 28670-1-AP, Proteintech ), ERBB2( 84046-1-RR, Proteintech ), and Ki67 ( 27309-1-AP, Proteintech ).

Techniques: Enzyme-linked Immunosorbent Assay, Knockdown, Sonication, Staining, Fluorescence, Microscopy, Immunohistochemistry, Expressing, Over Expression, Western Blot, Colony Assay, Viability Assay, Apoptosis Assay, In Vitro, Derivative Assay, Standard Deviation, Control

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A GSEA . The analysis shows a significant enrichment with a high NES of 2.0577, a P-value less than 0.001, and an FDR of 0.0119, indicating a strong association between SLC7A5 and this signaling pathway. B Correlation Analysis. TCGA database analysis revealed a positive correlation between ERBB2 and de novo lipid metabolism pathway-associated proteins ACLY, ACACA, and FASN, suggesting ERBB2 is involvement in lipid metabolism remodeling. C Proteasome Inhibition Assay. Knockdown of ERBB2 significantly reduces ACLY levels (line 2), which are restored upon treatment with 50 µM MG-132 for 6 h (line 4). D CHX Half-life Assay. ERBB2 knockdown shortens the half-life of ACLY (Left). Nuclear-cytoplasmic fractionation revealed that ERBB2 signaling selectively regulates ACLY activity/modification in the cytoplasmic compartment (Right). CHX. 10 µg/mL. E Denaturing IP Assay. Overexpression of ERBB2 decreases ACLY ubiquitination. F Co-Immunoprecipitation (Co-IP) Assay. Overexpression of ERBB2 enhances binding between CUL3 and ERBB2, while reducing binding between CUL3 and KLHL25. All in vitro data are derived from at least three independent experiments. G Proteasome Inhibition Assay. SLC7A5 knockdown increases ACLY (line 2), but ERBB2 knockdown further decreases ACLY levels (line 3). This decrease is inhibited by 50 µM MG-132 for 6 h (line 6). H Denaturing IP Assay. ERBB2 knockdown increases ACLY ubiquitination (line 2), which is reduced upon additional knockdown of CUL3(line 3) or KLHL25 (line 4). I Proteasome Inhibition Assay. ERBB2 knockdown decreases ACLY (line 2), but this decrease is reversed by knockdown of CUL3(line 3) or KLHL25 (line 4). MG-132 treatment (50 µM for 6 h) restores ACLY expression (line 7, 8). J Denaturing IP Assay. SLC7A5 overexpression reduces ACLY ubiquitination (line 2), which is increased upon ERBB2 knockdown (line 3). K ELISA. ELISA measurements of Tri-Glycerides, Free Fatty Acids, and Total Cholesterol showed that ERBB2 knockdown reverses the lipid metabolism remodeling caused by SLC7A5 overexpression. L Lipid Droplet Staining. LD540 staining demonstrated a reduction in lipid droplet accumulation induced by SLC7A5 following ERBB2 knockdown. Error bars represent standard deviation. Statistical significance is denoted as follows. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment.

Article Snippet: Prior to the staining process, the sections were deparaffinized and rehydrated, followed by heat-induced epitope retrieval using citrate buffer at 121 °C for 30 min. To inhibit endogenous peroxidase activity, sections were treated with 0.3% hydrogen peroxide in methanol for 30 min. Non-specific binding was mitigated by blocking with 10% normal bovine serum, after which the sections were incubated overnight at 4 °C with rabbit polyclonal antibodies targeting SLC7A5( 28670-1-AP, Proteintech ), ERBB2( 84046-1-RR, Proteintech ), and Ki67 ( 27309-1-AP, Proteintech ).

Techniques: Inhibition, Knockdown, Fractionation, Activity Assay, Modification, Over Expression, Ubiquitin Proteomics, Co-Immunoprecipitation Assay, Binding Assay, In Vitro, Derivative Assay, Expressing, Enzyme-linked Immunosorbent Assay, Staining, Standard Deviation, Control

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A – C Cell Function Experiments . ERBB2 knockdown reverses olaparib resistance induced by SLC7A5 overexpression. A Clonogenic Assay. B Apoptosis Assay. C Cell Viability Assay. D , E Kaplan−Meier survival curves of nude mice implanted orthotopically with CAOV-3 cells. In a xenograft survival study conducted in nude mice, 5 million CAOV-3 cells were subcutaneously injected. D Mice were divided into three groups, NC + shCtrl, SLC7A5 + shCtrl, and shSLC7A5 + shERBB2, with 20 mice per group. Once the tumors reached a volume of 50 mm³, each group was randomly divided into control and treatment subgroups. The treatment subgroup received olaparib every other day for a total of eight doses at 25 mg/kg, while the control group received an equivalent volume of DMSO. In accordance with animal ethics, mice were euthanized when tumor volumes reached 1000 mm³, which was recorded as the endpoint for survival. The median survival times were as follows. 27.5 days for the (NC + shCtrl + DMSO) group, 15 days for the (SLC7A5 + shCtrl + DMSO) group, 30 days for the (SLC7A5 + shERBB2 + DMSO) group, 45 days for the (NC + shCtrl + Olaparib) group, 20 days for the (SLC7A5 + shCtrl + Olaparib) group, and 47.5 days for the (NC + shCtrl +Olaparib) group (n = 10). E The relative extension of survival was statistically analyzed following olaparib treatment. All the in vitro data were obtained from at least three independent experiments. Error bars represent standard deviation. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment. F Clonogenic Assay . The knockdown of ERBB2 inhibits the proliferative effects induced by the overexpression of SLC7A5. G – K In vivo experiments. Subcutaneous implantation of CAOV-3 cells showed that ERBB2 inhibits SLC7A5-mediated tumor promotion. G Tumor. H Tumor Volume(mm 3 ). I Tumor Weight (g). J Immunohistochemistry (IHC). K Western Blot Analysis. All in vitro data points are derived from at least three independent experiments. Error bars represent the standard deviation. Statistical significance is indicated by the following symbols. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment.

Article Snippet: Prior to the staining process, the sections were deparaffinized and rehydrated, followed by heat-induced epitope retrieval using citrate buffer at 121 °C for 30 min. To inhibit endogenous peroxidase activity, sections were treated with 0.3% hydrogen peroxide in methanol for 30 min. Non-specific binding was mitigated by blocking with 10% normal bovine serum, after which the sections were incubated overnight at 4 °C with rabbit polyclonal antibodies targeting SLC7A5( 28670-1-AP, Proteintech ), ERBB2( 84046-1-RR, Proteintech ), and Ki67 ( 27309-1-AP, Proteintech ).

Techniques: Cell Function Assay, Knockdown, Over Expression, Clonogenic Assay, Apoptosis Assay, Viability Assay, Injection, Control, In Vitro, Standard Deviation, In Vivo, Immunohistochemistry, Western Blot, Derivative Assay

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A RT-PCR and WB Analysis. The knockout of SLC7A5 significantly inhibited the expression of ERBB2. Line left. HEY, Line right. SKOV3. B RT-PCR Analysis. RT-PCR showed that ELK1 knockdown significantly inhibits SLC7A5-mediated ERBB2 transcription. C Western Blot Analysis. Western blot demonstrated that ELK1 knockdown reverses the increase in ERBB2 protein levels caused by SLC7A5. D Nuclear and Cytoplasmic Fractionation. Nuclear and cytoplasmic fractionation combined with Western blot showed that SLC7A5 overexpression significantly promotes the nuclear translocation of pELK1. E Co-Immunoprecipitation (Co-IP). Co-IP confirmed SLC7A5- ELK1 protein-protein interactions. F Dual Luciferase Reporter Assay. This assay showed that SLC7A5 enhances ELK1 binding to the ERBB2 transcript in a dose-dependent manner. H RT-PCR Analysis. RT-PCR demonstrated that TDE significantly inhibits SLC7A5-mediated ERBB2 protein expression. G Dual Luciferase Reporter Assay. This assay showed that TDE significantly inhibits ELK1 binding to the ERBB2 transcript in a dose-dependent manner. I Chromatin Immunoprecipitation (ChIP) Assay. ChIP analysis showed that TDE significantly inhibits ELK1 binding to the ERBB2 transcript. J Dual Luciferase Reporter Assay. This assay demonstrated that ELK1 binds to the ERBB2 promoter sequence “CCTTCCATC”. Left. Schematic of dual luciferase reporter site mutations. Right. Dual luciferase assay results. K – M Proliferation Assay. TDE significantly mitigates the Olaparib resistance induced by SLC7A5 overexpression. K Edu Assay. L Colony-formation assay. M Cell Viability Test. All in vitro data are derived from at least three independent experiments. Error bars represent standard deviation. Statistical significance is denoted as follows. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment.

Article Snippet: Prior to the staining process, the sections were deparaffinized and rehydrated, followed by heat-induced epitope retrieval using citrate buffer at 121 °C for 30 min. To inhibit endogenous peroxidase activity, sections were treated with 0.3% hydrogen peroxide in methanol for 30 min. Non-specific binding was mitigated by blocking with 10% normal bovine serum, after which the sections were incubated overnight at 4 °C with rabbit polyclonal antibodies targeting SLC7A5( 28670-1-AP, Proteintech ), ERBB2( 84046-1-RR, Proteintech ), and Ki67 ( 27309-1-AP, Proteintech ).

Techniques: Reverse Transcription Polymerase Chain Reaction, Knock-Out, Expressing, Knockdown, Western Blot, Fractionation, Over Expression, Translocation Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Protein-Protein interactions, Luciferase, Reporter Assay, Binding Assay, Chromatin Immunoprecipitation, Sequencing, Proliferation Assay, EdU Assay, Colony Assay, In Vitro, Derivative Assay, Standard Deviation, Control

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer.

Article Snippet: Prior to the staining process, the sections were deparaffinized and rehydrated, followed by heat-induced epitope retrieval using citrate buffer at 121 °C for 30 min. To inhibit endogenous peroxidase activity, sections were treated with 0.3% hydrogen peroxide in methanol for 30 min. Non-specific binding was mitigated by blocking with 10% normal bovine serum, after which the sections were incubated overnight at 4 °C with rabbit polyclonal antibodies targeting SLC7A5( 28670-1-AP, Proteintech ), ERBB2( 84046-1-RR, Proteintech ), and Ki67 ( 27309-1-AP, Proteintech ).

Techniques:

Journal: bioRxiv

Article Title: PSP-0119: Targeted IRAK4 Degradation as a Novel Therapeutic Strategy for FLT3-Mutant AML

doi: 10.1101/2025.09.30.679569

Figure Lengend Snippet: (A ): Analysis of the AML patient’s microarray database (source: http://vizome.org/aml2/expression_strat/ showed that IRAK4 mRNA is significantly overexpressed in initial diagnosis (p<0.0001, n=16 each), relapse (p<0.001, n=16 each), residual disease (p=0.031, n=16 each), residual (p=0.034, n=16 each), FLT-3-ITD positive (p<0.0001, n=16 each), prior MDS (p=0.03, n=16 each), prior MPN (p<0.0001, n=16 each) and MDS- MPN+ve patients (p<0.0001, n=16 each) compared to healthy pooled CD34+ cells. On the other hand, (B ): IRAK1 mRNA expression was not significantly upregulated among these AML conditions than healthy pooled CD34+ cells. ( C ): IRAK4 mRNA is significantly upregulated in T(15:17), Inv(16)/t(16:16) and T(11q23)/MLL AML patients (source: Bloodspot.org ). ( D ): Kaplan-Meier analysis showed that IRAK4 mRNA overexpressor patients face early and greater proportions of deaths compared to low expressor patients (Source: GEPIA, http://gepia.cancer-pku.cn/detail.php?gene=IRAK4&clicktag=survival ). ( E ): IRAK1 mRNA is significantly upregulated in complex and T(11q23)/MLL karyotype presenting AML patients (source: Bloodspot.org ). ( F ): Kaplan-Meier analysis showed that IRAK1-mRNA overexpressor patients face early and great proportions of deaths than low expressor patients (Source: GEPIA, http://gepia.cancer-pku.cn/detail.php?gene=IRAK1&clicktag=survival ).

Article Snippet: The PVDF membrane containing the transferred protein was blocked in 5% fat free milk solution for 30 minutes and probed with rabbit IRAK4 antibody (Cell Signaling Tech, cat. no:4363, dilution: 1:1000), rabbit IRAK1 antibody (Cell Signaling Tech, cat. no:4504, dilution: 1:1000).

Techniques: Microarray, Expressing, Biomarker Discovery

Journal: bioRxiv

Article Title: PSP-0119: Targeted IRAK4 Degradation as a Novel Therapeutic Strategy for FLT3-Mutant AML

doi: 10.1101/2025.09.30.679569

Figure Lengend Snippet: In silico docking of PSP-0119 and PSP-0102 with CRBN and IRAK1 and IRAK4 complex. (A): In silico docking showed how glutarimide ring of lenalidomide interacts with CRBN and how pyrazolyl-pyridinyl portion engages with IRAK4. Red arrows point to both interactions. (B): Individual CRBN and IRAK4 residues interacting with PSP-0119 atoms are shown. (C): In silico docking showed how glutarimide ring of lenalidomide interacts with CRBN and how pyrazolyl-pyridinyl portion engages with IRAK1. (D): Individual CRBN and IRAK1 residues interacting with PSP-0102 atoms are shown.

Article Snippet: The PVDF membrane containing the transferred protein was blocked in 5% fat free milk solution for 30 minutes and probed with rabbit IRAK4 antibody (Cell Signaling Tech, cat. no:4363, dilution: 1:1000), rabbit IRAK1 antibody (Cell Signaling Tech, cat. no:4504, dilution: 1:1000).

Techniques: In Silico

Journal: bioRxiv

Article Title: PSP-0119: Targeted IRAK4 Degradation as a Novel Therapeutic Strategy for FLT3-Mutant AML

doi: 10.1101/2025.09.30.679569

Figure Lengend Snippet: ( A ): Hotspot kinase assay shows that despite drastic modifications on UR241-2, the kinase inhibition activity of PSP-0102 and PSP-0119 was not lost and showed IRAK4 kinase inhibition at 2.33nM. ( B ): PSP-0119 inhibited NF-kβ reporter activity induced by IL1β. ( C ): PSP-0119 pretreatment blocked the IL1β induced IRAK4 phosphorylation.

Article Snippet: The PVDF membrane containing the transferred protein was blocked in 5% fat free milk solution for 30 minutes and probed with rabbit IRAK4 antibody (Cell Signaling Tech, cat. no:4363, dilution: 1:1000), rabbit IRAK1 antibody (Cell Signaling Tech, cat. no:4504, dilution: 1:1000).

Techniques: Kinase Assay, Inhibition, Activity Assay, Phospho-proteomics

Journal: bioRxiv

Article Title: PSP-0119: Targeted IRAK4 Degradation as a Novel Therapeutic Strategy for FLT3-Mutant AML

doi: 10.1101/2025.09.30.679569

Figure Lengend Snippet: Repeated experiments showed PSP-0119 treatment did not degrade IRAK4 in THP-1 (FLT-3 wild-type) ( A ), FLT-3 wild-type AML patient ( B ) and normal bone marrow cells ( C ). In immunoblotting of FLT-3 normal AML patients ( C) , total ERK1/2 was used as a loading control as Actin and GAPDH showed variabilities. ( D ): PSP-0119 treatment degraded IRAK4 in MOLM-13 and MV-4-11 ( E ) FLT-3 mutant AML cells between doses 20-40nM during 24 hrs. of treatment ( D and E ). Normalized (IRAK4/GAPDH) pixel density is written in numbers in between the bands. ( F ): IRAK4/GAPDH normalized curves and calculated DC 50 of PSP-0119 against MV-4-11 and MOLM-13 are shown. ( G ): PS-0119 did not degrade IRAK1 in MOLM-13 cells. ( H ): PS-0119 strongly degraded IRAK1 in MV-4-11 cells.

Article Snippet: The PVDF membrane containing the transferred protein was blocked in 5% fat free milk solution for 30 minutes and probed with rabbit IRAK4 antibody (Cell Signaling Tech, cat. no:4363, dilution: 1:1000), rabbit IRAK1 antibody (Cell Signaling Tech, cat. no:4504, dilution: 1:1000).

Techniques: Western Blot, Control, Mutagenesis

Journal: bioRxiv

Article Title: PSP-0119: Targeted IRAK4 Degradation as a Novel Therapeutic Strategy for FLT3-Mutant AML

doi: 10.1101/2025.09.30.679569

Figure Lengend Snippet: ( A-B ): Bulk-sequencing of PSP-0119 treated MOLM-13 cells have identified gene affected by IRAK4 degradation. ( C ): Volcano plot of the upregulated and downregulated gene after IRAK1/4 degradation in MOLM-13 cells treated with PSP-0119 compared to vehicle. PS-0119 treatment led to downregulation of metabolic driver ENOS-1 gene in MOM-13 cells. Analysis of AML patient’s microarray data available at R2-Genomics and Visualization platform showed that ENOS-1 overexpression predicted poor prognoses in AML patients.

Article Snippet: The PVDF membrane containing the transferred protein was blocked in 5% fat free milk solution for 30 minutes and probed with rabbit IRAK4 antibody (Cell Signaling Tech, cat. no:4363, dilution: 1:1000), rabbit IRAK1 antibody (Cell Signaling Tech, cat. no:4504, dilution: 1:1000).

Techniques: Sequencing, Microarray, Over Expression

Journal: Cell Reports

Article Title: p53 Loss in Breast Cancer Leads to Myc Activation, Increased Cell Plasticity, and Expression of a Mitotic Signature with Prognostic Value

doi: 10.1016/j.celrep.2018.12.071

Figure Lengend Snippet:

Article Snippet: Primary antibodies used were: a rabbit monoclonal c-Myc antibody (1:250 in blocking solution, clone Y69, ab32072 Abcam); a rat monoclonal anti-CD49f (1:250 in blocking solution, clone GoH3, #555734, BD); a monoclonal mouse anti-Numb antibody (clone AB21) ( ).

Techniques: Derivative Assay, Recombinant, Flow Cytometry, Labeling, Expressing, Microarray, Software

Journal: Biology of reproduction

Article Title: Lin28a is dormant, functional, and dispensable during mouse oocyte-to-embryo transition.

doi: 10.1095/biolreprod.114.118703

Figure Lengend Snippet: FIG. 1. Lin28a and Lin28b expression analysis. A) Microarray data from the BioGPS database (NCBI accession number GSE1133 [36, 37]) show that Lin28a and Lin28b are highly expressed in the oocyte but not in somatic tissues. Data for each tissue represent relative average (n ¼ 2) fluorescence intensity of gene-detecting probes on the GNF1M platform (Affymetrix). Data from microarrays were normalized using the GC content adjusted Robust Multi-Array (gcRMA) algorithm and global median scaling, where the median was set to 200. B) Temporal expression profile of Lin28a transcripts during early development. Relative abundance of Lin28a transcripts was determined by real-time PCR. Expression data (CtDD values) obtained from individual

Article Snippet: The cells were then permeabilized for 15 min in PBS containing 0.1% Triton X-100, blocked in PBS containing 0.2% immunoglobulin G (IgG)-free BSA and 0.01% Tween-20 for 30 min (blocking solution), and then incubated with rabbit anti-LIN28 antibody (clone D1A1A; diluted 1:400 in blocking solution; Cell Signaling Technology) at 48C overnight.

Techniques: Expressing, Microarray, Fluorescence, Real-time Polymerase Chain Reaction

Journal: Biology of reproduction

Article Title: Lin28a is dormant, functional, and dispensable during mouse oocyte-to-embryo transition.

doi: 10.1095/biolreprod.114.118703

Figure Lengend Snippet: FIG. 2. Down-regulation of Lin28a and Lin28b transcripts by transgenic RNAi. A) Design of the transgenic RNAi vector pZP3EGFP-Lin28IR. Lin28a and Lin28b sequences used to produce the inverted repeat originated from 30UTR regions (Supplemental Fig. S1). Detailed description of the vector construction is described in Materials and Methods. B) Schematic depiction of simultaneous targeting of Lin28a and Lin28b by transgenic RNAi. C) Transgenic mice carrying the pZP3EGFP-Lin28IR transgene show uniform enhanced green fluorescent green protein (EGFP) expression in oocytes. A population of fully grown GV oocytes isolated from ovaries of a transgenic animal is shown, as seen with inverted fluorescence microscopy. Bar¼100 lm. D) Oocytes from mice carrying the pZP3EGFP-Lin28IR transgene show strong down-regulation of Lin28a and Lin28b expression. RT-PCR analysis was carried out as described in Materials and Methods. PCR-amplified amplicons after 45 cycles are shown.

Article Snippet: The cells were then permeabilized for 15 min in PBS containing 0.1% Triton X-100, blocked in PBS containing 0.2% immunoglobulin G (IgG)-free BSA and 0.01% Tween-20 for 30 min (blocking solution), and then incubated with rabbit anti-LIN28 antibody (clone D1A1A; diluted 1:400 in blocking solution; Cell Signaling Technology) at 48C overnight.

Techniques: Transgenic Assay, Plasmid Preparation, Expressing, Isolation, Fluorescence, Microscopy, Reverse Transcription Polymerase Chain Reaction, Amplification

Journal: Biology of reproduction

Article Title: Lin28a is dormant, functional, and dispensable during mouse oocyte-to-embryo transition.

doi: 10.1095/biolreprod.114.118703

Figure Lengend Snippet: FIG. 3. Transgenic RNAi prevents accumulation of LIN28A during oocyte-to-zygote transition. GV oocytes (GV), 1-cell embryos (1-cell), and 2-cell embryos (2-cell) from wild-type and transgenic females were stained with a-LIN28A antibody (red signal). DNA was stained with DAPI (blue signal). Bar ¼ 10 lm.

Article Snippet: The cells were then permeabilized for 15 min in PBS containing 0.1% Triton X-100, blocked in PBS containing 0.2% immunoglobulin G (IgG)-free BSA and 0.01% Tween-20 for 30 min (blocking solution), and then incubated with rabbit anti-LIN28 antibody (clone D1A1A; diluted 1:400 in blocking solution; Cell Signaling Technology) at 48C overnight.

Techniques: Transgenic Assay, Staining

Journal: Biology of reproduction

Article Title: Lin28a is dormant, functional, and dispensable during mouse oocyte-to-embryo transition.

doi: 10.1095/biolreprod.114.118703

Figure Lengend Snippet: FIG. 6. Model of Lin28 function during OET. The model integrates published data and results presented in this work. The maternal pool of Lin28a transcripts was translated during meiotic maturation and after fertilization (our data and those in ref. [39]). LIN28 does not target the mature Let-7 micro-RNA, but the precursor miRNA (pre-miRNA) released from the primary miRNA (pri-miRNA) transcript [26–30] suppresses zygotic expression of Let-7 (our data). The primary miRNA transcript (pri- miRNA) carrying miR-290-295 becomes expressed during the zygotic genome activation at the 2-cell stage, and miR-290-295 accumulate during early development [13, 54]. Superimposed on specific suppression of Let-7 is the global suppression of miRNA activity, which reduces the ability of mature miRNAs loaded on effector complexes to target imperfectly complementary target mRNAs [19, 20]. Likewise, oocytes and zygotes completely lacking canonical miRNAs (via maternal and maternal zygotic knock-out of Dgcr8, an essential miRNA biogenesis factor) develop to the blastocyst stage [20], further demonstrating that miRNAs are not important regulators of OET. Dynamics of P-bodies suggest that suppression of miRNA activity begins with the onset of oocyte growth and that miRNAs regain their function at approximately the 8-cell or morula stage [47].

Article Snippet: The cells were then permeabilized for 15 min in PBS containing 0.1% Triton X-100, blocked in PBS containing 0.2% immunoglobulin G (IgG)-free BSA and 0.01% Tween-20 for 30 min (blocking solution), and then incubated with rabbit anti-LIN28 antibody (clone D1A1A; diluted 1:400 in blocking solution; Cell Signaling Technology) at 48C overnight.

Techniques: Expressing, Activation Assay, Activity Assay, Knock-Out

Journal: Cell

Article Title: Mapping Systemic Inflammation and Antibody Responses in Multisystem Inflammatory Syndrome in Children (MIS-C)

doi: 10.1016/j.cell.2020.09.034

Figure Lengend Snippet:

Article Snippet: Bovin serum albumin , MP Biomedicals , Cat No. 160069.

Techniques: Recombinant, Blocking Assay, Staining, Microarray, Software, Immunoprecipitation