Journal: Journal of cellular signaling

Article Title: Chronic IL-1 Exposed AR + PCa Cell Lines Show Conserved Loss of IL-1 Sensitivity and Evolve Both Conserved and Unique Differential Gene Expression Profiles

doi:

Figure Lengend Snippet: Parental MDA-PCa-2b (MDA2b) and chronic IL-1 subline cells (MDA-αs1, MDA-αs2, MDA-βs1, MDA-βs2) were treated acutely for 3 days (A, C) or 6 days (B) with vehicle control (V), 25 ng/ml IL-1α (a), or 25 ng/ml IL-1β (b) and analyzed for cell viability using MTT (A, B) or protein accumulation by western blot (C). Acute IL-1 exposure reduces cell viability and proliferation, reduces full-length PARP (indicative of cell death activation), induces SOD2 and LCN2 protein accumulation (canonical IL-1-induced genes), and reduces AR and NKX3.1 (canonical AR target gene) protein accumulation in MDA-PCa-2b parental cells, but has little to no effect on the chronic IL-1 sublines. Thus, the IL-1 sublines evolved insensitivity to IL-1. Error bars, ± STDEV of 4 biological replicates; p-value, *≤ 0.05, **≤ 0.005, ***≤ 0.005, NS = not significant. Fold MTT optical density (OD) is normalized to treatment control. β-actin is the western blot loading control.

Article Snippet: MDA-PCa-2b cells were maintained in HPC1/20% FB Essence (FBE) containing 0.5 ng/ml IL-1α (Gold Bio, St. Louis, MO; 1110–01A-10) or IL-1β (Gold Bio, St. Louis, MO; 1110–01B-10) for approximately 4 months; and the proliferative colonies that emerged were expanded and termed MDA-PCa-2b IL-1α subline (MDA-αs) and MDA-PCa-2b IL-1β subline (MDA-βs).

Techniques: Western Blot, Activation Assay

Journal: Journal of cellular signaling

Article Title: Chronic IL-1 Exposed AR + PCa Cell Lines Show Conserved Loss of IL-1 Sensitivity and Evolve Both Conserved and Unique Differential Gene Expression Profiles

doi:

Figure Lengend Snippet: (A) MDA-PCA-2b parental (MDA2b) and chronic IL-1 subline (MDA-αs1, MDA-αs2, MDA-βs1, MDA-βs2) cells were treated acutely for 3 days with vehicle control, 25 ng/ml IL-1α, or 25 ng/ml IL-1β and analyzed by RT-qPCR for mRNA levels of the IL-1 receptor, IL-1R1 . Acute IL-1 exposure does not increase IL-1 receptor ( IL-1R1 ) mRNA levels in parental cells, suggesting basal IL-1R1 levels are sufficient to mediate IL-1 signaling. Furthermore, IL-1 does not show a differential effect on IL-1R1 mRNA levels in parental versus subline cells, suggesting subline insensitivity is independent of IL-1R1 levels. (B) Vehicle control treated cells were compared for basal mRNA levels of IL-1R1 and of canonical IL-1-induced genes, LCN2 , NOX1 , and SOD2 . IL-1R1, LCN2 , NOX1 , and SOD2 basal mRNA levels are comparable across the parental and subline cells, suggesting chronic IL-1 exposure does not induce constitutive activation of canonical IL-1 intracellular signaling. These data suggest that MDA-PCa-2b cell lines evolve insensitivity to exogenous chronic IL-1 exposure independent of IL-1R1 levels or constitutive activation of intracellular IL-1 signaling. Error bars, ± STDEV of 3 biological replicates; p-value, *≤ 0.05, **≤ 0.005, ***≤ 0.005, NS = not significant. For IL-1-treated cells, mRNA levels are normalized to vehicle control for each cell line. For basal expression, mRNA levels are normalized to the parental cell line.

Article Snippet: MDA-PCa-2b cells were maintained in HPC1/20% FB Essence (FBE) containing 0.5 ng/ml IL-1α (Gold Bio, St. Louis, MO; 1110–01A-10) or IL-1β (Gold Bio, St. Louis, MO; 1110–01B-10) for approximately 4 months; and the proliferative colonies that emerged were expanded and termed MDA-PCa-2b IL-1α subline (MDA-αs) and MDA-PCa-2b IL-1β subline (MDA-βs).

Techniques: Quantitative RT-PCR, Activation Assay, Expressing

Journal: Journal of cellular signaling

Article Title: Chronic IL-1 Exposed AR + PCa Cell Lines Show Conserved Loss of IL-1 Sensitivity and Evolve Both Conserved and Unique Differential Gene Expression Profiles

doi:

Figure Lengend Snippet: Parental (MDA-PCA-2b (MDA2b), LNCaP) and subline (MDA-αs1, MDA-αs2, MDA-βs1, MDA-βs2, LNas1, LNbs1) cells were treated acutely for 3 days with vehicle control, 25 ng/ml IL-1α, or 25 ng/ml IL-1β and analyzed by RT-qPCR for mRNA levels (A, B, C). Acute IL-1 exposure increases LCN2 , NOX1 , and SOD2 mRNA levels in parental MDA-PCa-2b and LNCaP cells, but acute IL-1 exposure has attenuated or no effect on mRNA levels in the subline cells. Thus, both LNCaP and MDA-PCa-2b cell lines show conserved intracellular response to acute IL-1-induced changes mRNA levels and evolve chronic IL-1 insensitivity independent of constitutive canonical IL-1 intracellular signaling. Error bars, ± STDEV of 3 biological replicates; p-value, *≤ 0.05, **≤ 0.005, ***≤ 0.005, NS = not significant. For IL-1-treated cells, mRNA levels are normalized to vehicle control for each cell line.

Article Snippet: MDA-PCa-2b cells were maintained in HPC1/20% FB Essence (FBE) containing 0.5 ng/ml IL-1α (Gold Bio, St. Louis, MO; 1110–01A-10) or IL-1β (Gold Bio, St. Louis, MO; 1110–01B-10) for approximately 4 months; and the proliferative colonies that emerged were expanded and termed MDA-PCa-2b IL-1α subline (MDA-αs) and MDA-PCa-2b IL-1β subline (MDA-βs).

Techniques: Quantitative RT-PCR

Journal: Cell & Bioscience

Article Title: The mechanisms by which hypothalamic neuroinflammation induced by neonatal cerebral ischemia–hypoxia leads to decreased thymic function via the HPA axis

doi: 10.1186/s13578-026-01543-w

Figure Lengend Snippet: HI induced the activations of TLR4/HMGB1/NF-κB signaling and microglia, leading to neuroinflammation in hypothalamus. A – B TNF-α and IL-1β on days 1, 3, 7, 14 and 21 after HI, n = 5–8. C Double immunofluorescence staining of Iba-1 and CD68. Scale bar = 20 μm. D Number of Iba-1 positive cells ( n = 4). E CD68 + / Iba-1 + cells rate ( n = 4). F – J CD11b, HMGB1, TLR4, p-P65-NF-κB and P65-NF-κB protein levels on day 7 after HI, n = 5. Data were expressed as the mean ± SD. Comparisons between the two groups were made using an unpaired T-test, * P < 0.05, ** P < 0.01, *** P < 0.001 versus Sham group.

Article Snippet: Serum hormone levels, along with hypothalamic tissue hormone and cytokine levels, were measured using CRH, ACTH, and CORT ELISA kits (Omnimabs, USA), as well as TNF-α and IL-1β ELISA kits (Proteintech, China), following the manufacturers’ instructions.

Techniques: Double Immunofluorescence Staining

Journal: Biology of Sex Differences

Article Title: Age and sex dependent effects of early overnutrition on metabolic parameters and the role of neonatal androgens

doi: 10.1186/s13293-016-0079-5

Figure Lengend Snippet: The limit of detection and intra- and inter-assay coefficients (CV) of variation for the hormone and cytokine assays

Article Snippet: Assay-on-demand kits (Applied Biosystems) were used to assess the mRNA levels of leptin (Rn00565158_m1), adiponectin (Rn00595250_m1), IL6 (Rn01410330_m1), IL1β (Rn01336189_m1), and TNFα (Rn01525859_g1) quantitative real-time PCR according to the manufacturer’s protocol and analyzed in an ABI PRISM 7000 Sequence Detection System (Applied Biosystems).

Techniques: Inter Assay, Intra Assay

Journal: Biology of Sex Differences

Article Title: Age and sex dependent effects of early overnutrition on metabolic parameters and the role of neonatal androgens

doi: 10.1186/s13293-016-0079-5

Figure Lengend Snippet: Glycemia and serum hormone and cytokine levels at postnatal days (PND) 10, 21, 30, 50, 85, and 150 in male (M) and female (F) rats raised in litters (L) 4 or 12 pups

Article Snippet: Assay-on-demand kits (Applied Biosystems) were used to assess the mRNA levels of leptin (Rn00565158_m1), adiponectin (Rn00595250_m1), IL6 (Rn01410330_m1), IL1β (Rn01336189_m1), and TNFα (Rn01525859_g1) quantitative real-time PCR according to the manufacturer’s protocol and analyzed in an ABI PRISM 7000 Sequence Detection System (Applied Biosystems).

Techniques:

Journal: Biology of Sex Differences

Article Title: Age and sex dependent effects of early overnutrition on metabolic parameters and the role of neonatal androgens

doi: 10.1186/s13293-016-0079-5

Figure Lengend Snippet: Relative mRNA levels of cytokines in inguinal and perigonadal adipose tissue at postnatal days (PND) 10, 21, 30, 50, 85, and 150 in male (M) and female (F) rats raised in litters (L) 4 or 12 pups

Article Snippet: Assay-on-demand kits (Applied Biosystems) were used to assess the mRNA levels of leptin (Rn00565158_m1), adiponectin (Rn00595250_m1), IL6 (Rn01410330_m1), IL1β (Rn01336189_m1), and TNFα (Rn01525859_g1) quantitative real-time PCR according to the manufacturer’s protocol and analyzed in an ABI PRISM 7000 Sequence Detection System (Applied Biosystems).

Techniques:

Journal: Biology of Sex Differences

Article Title: Age and sex dependent effects of early overnutrition on metabolic parameters and the role of neonatal androgens

doi: 10.1186/s13293-016-0079-5

Figure Lengend Snippet: Metabolic changes in males, females, and females that were androgenized (AF) on postnatal day (PND) 1. At PND10, there were significant differences in a the percentage of inguinal adipose tissue (IngAT) and serum b leptin, c adiponectin, and d interleukin (IL) 6 levels. At PND90, there were significant differences in e body weight, f glycemia and serum, g insulin, h leptin and i IL1β levels. The mRNA levels of j leptin, k adiponectin, l IL6, and m TNFα in perigonadal adipose tissue at PND90 were also affected. # p < 0.0001, ## p < 0.002, ### p < 0.0002, * p < 0.01, *** p < 0.005, @ p < 0.05. PND10: inguinal adipose tissue: N = 10; serum leptin and IL6 levels: N = 6; serum adiponectin: M and F: N = 8, AF: N = 9. PND90: body weight and glycemia: M: N = 8, F: N = 12, AF: N = 11; serum insulin, leptin, and Il1β levels: N = 6; mRNA levels for leptin, adiponectin, IL6, and TNFα: N = 6

Article Snippet: Assay-on-demand kits (Applied Biosystems) were used to assess the mRNA levels of leptin (Rn00565158_m1), adiponectin (Rn00595250_m1), IL6 (Rn01410330_m1), IL1β (Rn01336189_m1), and TNFα (Rn01525859_g1) quantitative real-time PCR according to the manufacturer’s protocol and analyzed in an ABI PRISM 7000 Sequence Detection System (Applied Biosystems).

Techniques: