Journal: International Journal of Molecular Sciences

Article Title: Murine Oncostatin M Has Opposing Effects on the Proliferation of OP9 Bone Marrow Stromal Cells and NIH/3T3 Fibroblasts Signaling through the OSMR

doi: 10.3390/ijms222111649

Figure Lengend Snippet: OP9 and NIH/3T3 cells show differential levels of OSMR and LIFR. ( A ) OP9 and NIH/3T3 cells were examined for LIFR and OSMR expression. BaF3 cells were used as a negative control. Protein molecular weight is labeled in kDa. Relative protein quantities are marked in red. ( B ) Microarray analysis of OP9 and NIH/3T3 cells showing Osmr and Lifr expression. The p -values were calculated by Transcriptome Analysis Console software. *** p < 0.001. ns = not significant. ( C ) OP9 and NIH/3T3 cells were analyzed for LIFR surface expression using FACS. ( D ) OP9 and NIH/3T3 cells were examined for expression of LIFR and OSMR in relation to different incubation periods with 10 ng/mL mOSM. Protein molecular weight is labeled in kDa. ( E ) OP9 and NIH/3T3 cells were examined for LIFR surface expression in relation to different incubation periods with 10 ng/mL mOSM. p -Values were calculated using one-way ANOVA and Bonferroni post-test. * p <0.05 and *** p < 0.001. ns = not significant.

Article Snippet: Murine wildtype Lifr cDNA (MR221059) was obtained from OriGene (Rockville, MD, USA).

Techniques: Expressing, Negative Control, Molecular Weight, Labeling, Microarray, Software, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Murine Oncostatin M Has Opposing Effects on the Proliferation of OP9 Bone Marrow Stromal Cells and NIH/3T3 Fibroblasts Signaling through the OSMR

doi: 10.3390/ijms222111649

Figure Lengend Snippet: OSMR downregulation attenuates OSM effects on proliferation. ( A , B ) Immunoblot quantifying the shRNA-induced knockdown of Osmr (n = 4) and Lifr (n = 5) in ( A ) OP9 and ( B ) NIH/3T3 cells. The shRNAs Osmr 5 and Lifr 1 were used in subsequent experiments. Protein molecular weight is labeled in kDa. Relative protein quantities are indicated in red. ( C ) Relative quantification of S phase of OP9 and NIH/3T3 cells carrying a knockdown for either Osmr , Lifr, or a Renilla control in presence or absence of 10 ng/mL mOSM. Cell cycle analysis was performed as described before. One-way ANOVA and Bonferroni post-test. *** p < 0.001. ns = not significant. ( D ) Immunoblot quantifying the LIFR overexpression in OP9 and NIH/3T3 cells. Protein molecular weight is labeled in kDa. Relative protein quantities are indicated in red. ( E ) Lifr overexpressing OP9 and NIH/3T3 cells were examined for LIFR surface expression compared to cells carrying the mock control. ( F ) Relative quantification of S phase of OP9 and NIH/3T3 cells transduced with Lifr or empty vector (MiG). Cell cycle analysis was performed as described before. One-way ANOVA and Bonferroni post-test. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Murine wildtype Lifr cDNA (MR221059) was obtained from OriGene (Rockville, MD, USA).

Techniques: Western Blot, shRNA, Knockdown, Molecular Weight, Labeling, Quantitative Proteomics, Control, Cell Cycle Assay, Over Expression, Expressing, Transduction, Plasmid Preparation

Journal: bioRxiv

Article Title: Targeting CREB remodels the immune microenvironment to enhance immunotherapy responses in pancreatic cancer

doi: 10.64898/2025.12.04.691935

Figure Lengend Snippet: Cancer cell intrinsic CREB transcriptionally regulates LIF expression, driving macrophage-modulatory signaling in PDAC. (A) Experimental schematic demonstrating RNA sequencing (RNA-seq) analysis conducted in KPC mouse CREB WT and CREB KO tumor cells. (B) Bubble plot visualizing differentially regulated pathways (using molecular signature databases) in CREB KO transcriptomics compared with CREB WT tumor cells (n=3). (C) Ridge plot showing normalized enrichment scores for the hallmark JAK–STAT3 signaling pathway across different cellular constituents in the human HTAN pancreatic cancer dataset from treatment-naïve patients. (D) Western blot image depicting pSTAT3 activation, along with tSTAT3, vinculin and vimentin in mouse RAW macrophage 264.7 cell line treated with either recombinant (r) LIF alone (1 µg/mL) or in combination with EC359 (LIFR blockade, 0.2µM). (E) Chromatin Immunoprecipitation sequencing (ChIP-seq) peak signals visualized as heat maps which depict CREB binding sites across genomic regions in KPC mouse pancreatic tumor cells. The adjacent call out boxes with the heat maps showing essential genes regulated via CREB as its downstream mediators. (F) Integrative Genome Viewer (IGV) plot visualizing occupancy of CREB binding peaks in ChIP-seq data at the site of mouse Lif promoter gene regulatory sequences. (G-H) Violin dot plots showing downregulation of LIF expression via qPCR and ELISA in bulk tumor lysates of CREB KO KPC orthotopic PDAC tumors as compared to CREB WT with n=3-7 mice per group. (I) Representative photomicrographs of whole pancreas along with its corresponding quantification depicting significantly reduced LIF expression via IHC in age matched KPC C -/- as compared to KPC mice. Individual data points with mean ± SEM are shown and compared by two-tailed unpaired t test. *p<0.05; **p<0.01; ns non-significant (p>0.05).

Article Snippet: Conditioned medium was collected from cells, cleared by centrifugation, and analyzed using a commercially available sandwich ELISA kit for mouse LIF (# MLF00; R&D systems), mouse IFN-ψ (# MIF00-1; R&D systems) according to the manufacturer’s instructions.

Techniques: Expressing, RNA Sequencing, Western Blot, Activation Assay, Recombinant, ChIP-sequencing, Binding Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test