Fig 1: Regulation of Ccnd2 expression by prolactin signaling in β-cells.(A, B) mRNA expression of the indicated molecules in INS-1 cells transfected with siRNA targeting MafA (A) or Prlr (B) relative to INS-1 cells transfected with control siRNA. n = 4 for both (A) and (B). (C–P) INS-1 cells were transfected with control siRNA or siRNA targeting Prlr (si-Prlr) or MafA (si-MafA). At 48 hours after transfection, the cells were cultured in defined serum-free medium for 24 hours for serum deprivation, followed by the addition of prolactin (PRL) or PBS as a control. (C, D) The immunoprecipitates obtained with anti-Stat5B antibody from 1% TX-100 soluble fractions of INS-1 cells transfected with control si-RNA, si-Prlr or si-MafA, and with or without PRL stimulation were immunoblotted with either anti-Stat5B (C) or anti-phosphotyrosine antibody (D). n = 3. (E, F) 1% TX-100 soluble fractions of INS-1 cell lysates were immunoblotted with either anti-Stat5B (E) or anti-phosphorylated Stat5 antibody (F). n = 3. (G) Dose-dependent effects of PRL on the phosphorylation of Stat5. n = 2. (H–K) Localization of Stat5B (red), β-catenin (green) and nuclei, which were counterstained with DAPI (blue), in INS-1 cells that were transfected or not with si-Prlr and stimulated or not with PRL. n = 3. (L–N) Localization of Stat5B (red), β-catenin (green) and nuclei, which were counterstained with DAPI (blue), in INS-1 cells with or without PRL stimulation and with or without exposure to AG490, a Jak2 inhibitor. n = 3. (O, P) The amount of phosphorylated Stat5, Ccnd2 and Lamin A/C (control) in the nuclear fraction of INS-1 cells with (w/) or without (w/o) PRL stimulation after serum deprivation (O), and quantification of the results (P). n = 4. (Q) mRNA expression of the indicated molecules in INS-1 cells transfected with siRNA targeting Prlr relative to INS-1 cells transfected with control siRNA. n = 3 for each molecule. The data represent the mean ± S.E.M. in (A), (B), (P) and (Q), and representative results are shown in (C–O). Asterisks means p<0.05.
Fig 2: Effect of PRL in vitro on alveolar macrophages (cell line MHS). a Percentage of infected cells and the number of phagocytosed bacilli per cell after 1 h of incubation with M. tuberculosis in the presence of PRL before, during, and after bacterial exposure. Preincubation with PRL is the most effective method to induce phagocytosis. b Determination of intracellular live bacilli in macrophages infected after 1 h and 7 days, with or without PRL. There is significant intracellular bacteriolysis after 7 days of incubation with PRL. c Determination of superoxide and NO (d) in infected and noninfected alveolar macrophages incubated with PRL (two-way ANOVA with Sidak multiple comparisons; data are expressed as mean ± SD. bp < 0.05. cp < 0.001. dp < 0.01, p < 0.001). Two independent experiments were performed with similar results.
Fig 3: Effect of ovine PRL treatment in mice with late progressive disease. a Survival curve of mice treated with PRL. Statistical significance was evaluated using the log-rank (Mantel-Cox) test (p = 0.6357). b Pulmonary bacillary load determined by CFUs; there is a significant reduction after 2 months of treatment. c Surface area affected by pneumonia assessed by automated morphometry. d Gene expression of TNFα and IFNγ determined by RT-PCR (data are expressed as mean ± SD. a log-rank (Mantel-Cox) test. b–d Two-way ANOVA with Sidak multiple comparisons. bp < 0.01).
Fig 4: Effect of PRL in vitro on pneumocytes type II (cell line A549). a Gene transcription of surfactant protein A, surfactant protein D (b), and human cathelicidin LL-37 (c) in the indicated experimental conditions after 1 and 21 h of incubation with PRL. d Intracellular bacilli load, determined by CFUs in pneumocytes type II after one and 21 h of infection, with or without PRL (data are expressed as mean ± SD; two-way ANOVA with Sidak multiple comparisons. d p < 0.05).
Fig 5: Kinetics of prolactin (PRL) gene expression during pulmonary tuberculosis. a Progressive increase in bacillary loads in the lungs of mice infected with M. tuberculosis at different post-infection time points. b PRL gene transcription determined by RT-PCR throughout the course of infection. Data are shown as mean ± SD (n = 5 mice per time point). c–f Representative micrographs show PRL detection by immunohistochemistry. c No PRL is detected in the lungs of control, non-infected mice. d In contrast, alveolar epithelial cells display strong PRL immunostaining after one day of M. tuberculosis infection. e After two weeks of infection, granulomas contain some lymphocytes and macrophages with positive PRL immunostaining. f After four months of infection, certain lymphocytes and activated macrophages—large cells with abundant, compact cytoplasm (arrowheads) in pneumonic areas—as well as bronchial epithelial cells (arrow), exhibit PRL immunoreactivity, while foamy macrophages are negative (asterisks).
Supplier Page from R&D Systems, a Bio-Techne Brand for Prolactin Protein
Available conjugates: Sizes Available: 50 ug