3t3 l1 murine preadipocyte cell line  (ATCC)

Journal: Cell Reports Medicine

Article Title: An aerosolized dual-action autotaxin inhibitor-PPARγ agonist for the treatment of pulmonary fibrosis

doi: 10.1016/j.xcrm.2026.102778

Figure Lengend Snippet: EL244 is a potent PPARγ agonist with therapeutic potential In (A–F), EL244 ability to differentiate 3T3-L1 fibroblasts to adipocytes in the presence of insulin was tested. (A) Oil red O staining of lipid droplets indicative of adipocyte differentiation. The cocktail containing insulin (Ins.), dexamethazone (DEX), and 3-isobutyl-1-methylxanthine (IBMX) was used as a positive control. Scale bars, 100 μm. (B) Absorbance of oil red O staining in cells ( n = 5, 5, 5, 5). (C–E) mRNA levels of PPARγ target genes Adiponectin (C), Fabp4 (D), and Cd36 (E) in 3T3-L1 or the differentiated adipocytes. mRNA expression was interrogated with RT-qPCR; values were normalized to the expression of B2m and presented as fold change over control ( n = 3, 5, 5, 5, 5). (F) mRNA levels of PPARγ in 3T3-L1 or the differentiated adipocytes. mRNA expression was interrogated with RT-qPCR; values were normalized to the expression of B2m and presented as fold change over control ( n = 3, 5, 5, 5, 5). In (G–R), C57bl/6 male mice were treated with high-fat diet (HFD) for 10 weeks and three injections of streptozotocin (STZ, 40mg/kg). In the last 2 weeks of the HFD treatment, a group of mice was treated daily with EL244 (50 mg/kg). (G) Weight curves of the different groups of mice during the HFD, STZ, and EL244 administration. (H) Oral glucose tolerance test (oGTT) after oral administration of a 10% glucose solution (1 g/kg). (I) Insulin resistance as assessed by the area under curve (AUC) of oGTT ( n = 5, 8, 5). (J) Glucose levels in the beginning of the oGTT ( n = 5, 8, 5). (K–R) mRNA levels of PPARγ and PPARγ target genes in the adipose tissue (K–N) and liver (O–R) from mice subjected to HFD, STZ, and EL244 treatment. mRNA expression was interrogated with RT-qPCR; values were normalized to the expression of B2m and presented as fold change over control. (K) Glut1 ( n = 5, 7, 4), (L) Fabp4 ( n = 5, 7, 4), (M) Cd36 ( n = 4, 8, 5), and (N) PPARγ ( n = 5, 5, 4) mRNA levels in adipose tissue. (O) Glut1 ( n = 5, 8, 6), (P) Fabp4 ( n = 5, 8, 6), (Q) Cd36 ( n = 5, 7, 6), and (R) PPARγ ( n = 5, 8, 6) mRNA levels in liver. Data in box and whiskers include the median (line), interquartile range (box), and minimum and maximum range (tails). Each dot represents a biological replicate. Following normality testing, statistical significance was assessed with one-way ANOVA and Tukey’s post-hoc test (B–D), (F), (I–L), (N), (O), (P), and (R) or Welch ANOVA and Games-Howell’s post-hoc test (E), (M), and (Q). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, respectively. See also .

Article Snippet: 3T3-L1 cell line , ATCC , N/A.

Techniques: Staining, Positive Control, Expressing, Quantitative RT-PCR, Control

Journal: Cells

Article Title: Punicic Acid in Ovarian Cancer: Anticancer Activity and Mechanistic Insights

doi: 10.3390/cells15090792

Figure Lengend Snippet: Differential Effects of Punicic Acid and α-Eleostearic Acid on OC and Normal Cell Lines. ( a ) HEYA8 OC cells and NFT epithelial cell lines (P201 and P211) were treated for 48 h with methanol control (CTL, black bars), 20 µM PunA (medium grey bars), 25 µM PunA (dark grey bars) or 20 µM cisplatin (CP, light grey bars). Data are presented as mean ± SD of six technical replicates, representative of three independent experiments. Statistical significance was determined by one-way ANOVA with Dunnett’s multiple comparisons test versus control. **** p < 0.0001, ns: not significant. ( b ) OC cells and normal cell lines were treated with increasing concentrations of PunA (0, 5, 10, 20, and 40 μM) for 48 h, and cell viability was determined by the MTS assay. Data represent mean ± SD from three independent experiments. Statistical analysis was performed using two-way ANOVA followed by Dunnett’s multiple comparisons test. No significant differences were observed at baseline (0 μM; all p > 0.9999). At 5 μM, HEYA8 cells showed significantly lower viability compared to P201 ( p = 0.0196), P211 ( p = 0.0002), E7 ( p = 0.0005), and 3T3-L1 ( p = 0.0002), with all comparisons reaching p < 0.0001 at higher concentrations. ( c ) Structural comparison of PunA (cis-9, trans-11, cis-13) and α-ESA (cis-9, trans-11, trans-13), both conjugated trienoic FFAs differing only in the π-bond orientation at the omega-5 carbon. ( d ) HEYA8 and P201 cells were treated for 48 h with varying concentrations of PunA and α-ESA. While α-ESA reduced viability in both HEYA8 and P201 cells, PunA showed a dose-dependent reduction in HEYA8 viability with minimal cytotoxicity in P201 cells. Data are presented as mean ± SD of six technical replicates, representative of three independent experiments. Statistical significance was determined by two-way ANOVA with Dunnett’s post hoc test versus control. * p < 0.05, **** p < 0.0001. ( e ) Summary table of PunA IC 50 values between normal cell lines (top four rows) and OC cell lines (bottom three rows).

Article Snippet: The mouse preadipocyte 3T3-L1 cell line (Cat # CL-173) was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Control, MTS Assay, Comparison

Journal: Cells

Article Title: Punicic Acid in Ovarian Cancer: Anticancer Activity and Mechanistic Insights

doi: 10.3390/cells15090792

Figure Lengend Snippet: Effects of PunA Alone and in Combination with Cisplatin on Cell Viability, Drug Interaction, Resistance, and Lipidic Microenvironment. ( a ) HEYA8 cells were treated with PunA (15 or 30 μM) for 48 h, and cell viability was measured using an MTS assay. Statistical significance was determined by one-way ANOVA with Dunnett’s multiple comparisons test versus control. ( b ) HEYA8 cells were treated with cisplatin (30 μM), PunA (30 μM), or their combination for 48 h, followed by MTS analysis. Statistical significance was determined using one-way ANOVA followed by Šídák’s multiple comparisons test. ( c ) Drug interaction between PunA and cisplatin was evaluated using the Highest Single Agent (HSA) model by comparing predicted and observed inhibition. Data ( a – c ) are presented as mean ± SD of six technical replicates, representative of two independent experiments. * p < 0.05, **** p < 0.0001, ns: not significant. ( d ) Cisplatin-sensitive and -resistant cell lines (41M/41McisR and A2780/A2780cisR) were treated with increasing concentrations of PunA for 48 h (0 μM–60 μM for 41M pair, upper panel; 0 μM–75 μM for A2780 pair, lower panel). Data are presented as mean ± SD of six technical replicates, representative of three independent experiments. Statistical significance was assessed by two-way ANOVA with Dunnett’s post hoc test. For 41M/41McisR, the interaction, PunA factor, and drug-resistance factor were all significant ( p < 0.0001). For A2780/A2780cisR, the interaction was significant ( p = 0.0341), as were the PunA factor ( p < 0.0001) and drug-resistant factor ( p = 0.0019). ( e ) HEYA8 cells were cultured in DF or UD 3T3-L1 conditioned medium and treated with increasing concentrations of PunA (0–100 μM) for 48 h. Cell viability was measured by MTS assay, normalized to untreated controls. Data are presented as mean ± SD of six technical replicates, representative of three independent experiments. Statistical significance was assessed by two-way ANOVA with Dunnett’s post hoc test. The interaction was significant ( p = 0.0003), along with the PunA factor ( p < 0.001) and DF conditioned medium factor ( p = 0.0008).

Article Snippet: The mouse preadipocyte 3T3-L1 cell line (Cat # CL-173) was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: MTS Assay, Control, Inhibition, Cell Culture