primary antibodies for lin28b  (Cell Signaling Technology Inc)

Journal: Cell reports

Article Title: Ribosomal protein control of hematopoietic stem cell transformation through regulation of metabolism

doi: 10.1016/j.celrep.2025.116688

Figure Lengend Snippet: (A) Leukemia explants from MLL-AF9-knockin Rpl22 +/+ (M82) and Rpl22 −/− (M109) mice were analyzed by RNA-seq. A heatmap of 2,671 differentially expressed genes is displayed. (B) Top 20 upregulated essential genes in Rpl22 −/− leukemias identified using the CRISPR DEPMAP CERES dataset across human leukemia cell lines and displayed as a bubble plot of the impact of their genetic disruption on leukemia survival. (C) Differentially expressed genes were subjected to pathway analysis (Protein Analysis Through Evolutionalary Relationships; PANTHER) with the top upregulated and downregulated pathways displayed. The number of differentially expressed genes per pathway is indicated.

(D) Bubble plot of the top five Reactome pathways depicting the most upregulated metabolic pathways in Rpl22 −/− MLL-AF9 leukemias. (E) qPCR analysis of Lin28b mRNA expression in Rpl22 +/+ and Rpl22 −/− MLL-AF9 leukemias. Statistical significance was calculated using a two-tailed t test with Welch’s correction. Normalized Lin28b expression is depicted graphically as the mean +/− SD. (F) Heatmap of 103 Lin28b gene targets induced in Rpl22 −/− MLL-AF9 leukemias, subdivided based on whether they are direct Lin28b targets or are indirectly regulated through Lin28b modulation of Let7 micro-RNAs (mIRs). (G) Lipid content of Rpl22 +/+ and Rpl22 −/− leukemias as measured by Nile red staining. Statistical significance was calculated using a two-tailed t test with Welch’s correction. Normalized lipid content is depicted graphically as mean +/− SD. (H) Triacylglycerol (TG) levels were measured on detergent extracts of Rpl22 +/+ and Rpl22 −/− leukemias using a colorimetric assay quantifying oxidized glycerol liberated by lipase digestion. Mean ± SD of triplicate measures of nMol TG per ng of protein were expressed graphically. Statistical significance was determined using one-way ANOVA. (I) The effect of inhibiting TG synthesis using DGAT1 inhibitor (DGAT1-IN-1) at 25 μM on the growth of Rpl22 +/+ and Rpl22 −/− leukemias was assessed by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay. Triplicate measures were expressed graphically as mean ± SD for each different drug concentration. Statistical significance was determined by two-way ANOVA.

Article Snippet: Lin28b Taqman Primer/Probe set , ThermoFisher , Mm01190673_m1.

Techniques: Knock-In, RNA Sequencing, CRISPR, Disruption, Expressing, Two Tailed Test, Staining, Colorimetric Assay, MTT Assay, Concentration Assay

Journal: Advanced Science

Article Title: PARPi Combining Nanoparticle LIN28B siRNA for the Management of Malignant Ascites

doi: 10.1002/advs.202510547

Figure Lengend Snippet: Aberrant LIN28B expression and DNA repair pathway are associate with MSE. (a) Integrated analysis combining scRNA‐seq data from ascites in a constructed preclinical OC model; scRNA‐seq data from MA in OC patients ( n = 13), and gene mutation profiles from MSE in cancer patients ( n = 442) to explore novel dual‐targeting therapeutic strategies (created at BioRender.com). (b) Venn diagram analysis of DEG targets between the preclinical OC model and OC patients (Nat Cancer. 2023). GO‐BP enrichment analysis for (c) SNV gene mutations and (d) CNV gene mutations in MSE from cancer patients. (e) Waterfall plot for top 45 SNV genes and relative pathways. Genotype Quality (GQ): Minimum threshold of 20 (99% confidence in the genotype call). Read Depth (DP): Site‐specific depth filters (DP ≥ 10 for high‐confidence calls, adjusted for cohort mean depth). lternate Allele Support: Minimum alternative allele reads (≥ 3) and fraction (AF ≥ 0.25 for heterozygous germline calls). opulation Frequency: Filtering against population databases (gnomAD allele frequency < 0.01 for rare variants, < 0.001 for ultra‐rare). (f) Heatmap of CNV genes in MSE from cancer patients ( n = 442). Quality Metrics: Segment‐wise log2 ratio standard deviation or confidence score (e.g., CNVkit quality score > 0.5). Size Threshold: Typically, > 1 kb for targeted sequencing and > 10–50 kb for whole‐genome sequencing to exclude small, noisy calls. Copy State Threshold: Log2 ratio thresholds define copy states (e.g., deletion: log2 ratio < −0.4, duplication: log2 ratio > 0.2).

Article Snippet: The following antibodies were used: anti‐human LIN28B (Cell Signaling Technology, 4196S, 1:1000), anti‐mouse Lin28b (Proteintech, 11724‐1‐AP, 1:500), anti‐β‐actin (ABclonal, AC026, 1:10000), StarBright Blue 520 goat anti‐mouse IgG (Bio‐Rad, 12005867, 1:10000), Alexa Fluor Plus 800 goat anti‐rabbit IgG (H + L) (Invitrogen, A32735, 1:10000), Anti‐Mouse CD8a‐Purified In vivo (C375‐25 mg), Anti‐Mouse NK1.1‐Purified In vivo (N123‐25 mg), Human/Primate IL‐6 Mab(Clone 6708, MAB206‐SP, R&D), Human TNF‐alpha Mab(Clone28401, MAB610‐SP, R&D), anti‐CD31 antibody (Thermo Fisher Scientific, MA3105, 1:200), and anti‐TER‐119 antibody (BD Biosciences, 557915, 1:100).

Techniques: Expressing, Construct, Mutagenesis, Standard Deviation, Targeted Sequencing, Sequencing

Journal: Advanced Science

Article Title: PARPi Combining Nanoparticle LIN28B siRNA for the Management of Malignant Ascites

doi: 10.1002/advs.202510547

Figure Lengend Snippet: Synthesis and characterization of DSSP@lip‐PEG‐FA targeting LIN28B.(a) Schematic illustration of the synthesis process for siRNA/DSSP@lip‐PEG‐FA. Representative TEM images and size distribution profiles of siRNA/DSSP NPs (b) and siRNA/DSSP@lip‐PEG‐FA (c). (d) hydrogen peroxide (H 2 O 2 )‐ and glutathione (GSH)‐responsive siRNA release profiles under different conditions (pH 7.4, pH7.4& 1 mM H 2 O 2 , and pH 7.4 & 10 mM GSH) over 24 h. (e) Confocal microscopy images (scale bars: 50 µm) and quantitative analysis of cellular uptake efficiency for free siRNA, siRNA/DSSP@lip‐PEG, and siRNA/DSSP@lip‐PEG‐FA. (f) Flow cytometric analysis quantifying Cy3‐labeled siRNA fluorescence intensity in treated cells. (g) Flow cytometry evaluation of folate receptor (FA)‐targeting specificity. (h) Mean fluorescence intensity (MFI) comparison across different treatment groups at 6 h post‐incubation. (i) Confocal images showing intracellular distribution of siRNA/DSSP@lip‐PEG‐FA (scale bar: 10 µm). (j) Quantitative colocalization analysis performed using ImageJ software. (k) and (l) Western blot analysis of Lin28b protein expression in A2780 and ID8 ovarian cancer cells following various treatments. Data are presented as mean ± SD ( n = 3, unpaired t‐test). n = independent biological replicates. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant.

Article Snippet: The following antibodies were used: anti‐human LIN28B (Cell Signaling Technology, 4196S, 1:1000), anti‐mouse Lin28b (Proteintech, 11724‐1‐AP, 1:500), anti‐β‐actin (ABclonal, AC026, 1:10000), StarBright Blue 520 goat anti‐mouse IgG (Bio‐Rad, 12005867, 1:10000), Alexa Fluor Plus 800 goat anti‐rabbit IgG (H + L) (Invitrogen, A32735, 1:10000), Anti‐Mouse CD8a‐Purified In vivo (C375‐25 mg), Anti‐Mouse NK1.1‐Purified In vivo (N123‐25 mg), Human/Primate IL‐6 Mab(Clone 6708, MAB206‐SP, R&D), Human TNF‐alpha Mab(Clone28401, MAB610‐SP, R&D), anti‐CD31 antibody (Thermo Fisher Scientific, MA3105, 1:200), and anti‐TER‐119 antibody (BD Biosciences, 557915, 1:100).

Techniques: Confocal Microscopy, Labeling, Fluorescence, Flow Cytometry, Comparison, Incubation, Software, Western Blot, Expressing