Journal: mAbs

Article Title: Discovery and characterization of prolactin neutralizing monoclonal antibodies for the treatment of female-prevalent pain disorders.

doi: 10.1080/19420862.2023.2254676

Figure Lengend Snippet: Figure 3. Selectivity and cross-reactivity of PL 200,039. (a) hPRLR-expressing CHOK1SV GS-KO cells were stimulated for 15 minutes at 37°C with constant 20 nM hPRL, 20 nM hGH, or 350 nM hPL preincubated with varying concentrations (0–150 nM or 0–2.6 µM) of PL 200,039 for 30 minutes prior to stimulation. Data was compiled from three independent experiments and normalized to response of cells stimulated in the absence of antibody. Values represent mean ± SEM. (b) CHOK1SV GS-KO cells expressing hPRLR or mPRLR were stimulated as before with constant 20 nM nhpPRL or 20 nM rPRL, respectively, preincubated for 30 minutes with varying concentrations (0–150 nM) of PL 200,039 prior to stimulation. Data was compiled from two (rPRL) or three (nhpPRL) independent experiments and normalized to the response of cells stimulated in the absence of antibody. Values represent mean ± SEM.

Article Snippet: PRLR cell line development hPRLR (Origene, #RC209266) and mPRLR open reading frames were subcloned into a single gene GS expression vector (Lonza).

Techniques: Expressing

Journal: Cancer research

Article Title: Prolactin promotes fibrosis and pancreatic cancer progression

doi: 10.1158/0008-5472.CAN-18-3064

Figure Lengend Snippet: (A-D) Immunostaining against PRLR on tissues obtained from pancreatic cancer patients showed no PRLR expression in the normal pancreatic parenchyma (A), whereas it was expressed in high grade PanINs (B). PRLR could be expressed heterogeneously (C) or uniformly (D) in PDAC cells. Arrows in (C) highlight PRLR-negative PDAC cells. (E) Boxplot of PRLR gene expression stratified by class based on the study by Bailey et al. (34). (F) Kaplan-Meier analysis comparing survival of patients having either high or low PRLR expression (n=54, Median survival 27 vs 17 months). (G) PRLR was also expressed in subset of stromal cells (arrowheads). (H, I) Similar to their human counterparts, mouse PanIN (H) and PDAC (I) cells expressed PRLR. Arrows in (H) show a subset of PRLR-positive stromal cells in KC mice. N:normal pancreas, asterisks highlight early PanINs. Bars 20μm.

Article Snippet: PRLR expression was silenced with commercially obtained shRNA- containing lentiviral particles (Origene, Rockville MD).

Techniques: Immunostaining, Expressing, Gene Expression

Journal: Cancer research

Article Title: Prolactin promotes fibrosis and pancreatic cancer progression

doi: 10.1158/0008-5472.CAN-18-3064

Figure Lengend Snippet: (A, B) Immunostaining of tissues obtained from KC pancreas using antibodies against E-cadherin and F4/80 (A) or prolactin and F4/80 (B) showed expression prolactin in macrophages surrounding the PanIN lesions (dashed lines). Inset in (B) highlights prolactin-expressing macrophages. (C) Wild type BMD-macrophages were cultured with conditioned media collected from activated wild type or Hmgb1−/− platelets showed that prolactin expression in macrophages can be induced by HMGB1. (D) Immunostaining of tissues obtained from KCH pancreas using antibodies against prolactin and F4/80 showed lower expression of prolactin in macrophages surrounding the PanIN lesions (dashed lines). Inset in (D) highlights prolactin-expressing macrophages. (E) Representative flow cytometric analysis of PRLR expression on cells gated for CD11b and F4/80 expression showed no PRLR expression in bone marrow derived macrophages. (F-H) Representative flow cytometric analysis of CD11c, CD206 or PRLR expression on cells gated for CD11b and F4/80 collected from non-injured pancreas (F), on day 3 post-caerulein treatment (G), or KC mice (H) showed that PRLR+ macrophages could be exclusively found within CD11b+/F4/80+/CD11c+/CD206+ population. (I) Quantitative analysis of different macrophage populations in the non-injured adult pancreas, day 3 post-caerulein treated, or KC pancreas. Results in (I) are expressed as mean ± standard error of the mean (SEM) and analyzed statistically by Student’s t-test, *p<0.05, **p<0.01, ***p<0.001, ****p≤0.0001. (n=3 mice, and 3 replicates for each flow). Bars 20μm.

Article Snippet: PRLR expression was silenced with commercially obtained shRNA- containing lentiviral particles (Origene, Rockville MD).

Techniques: Immunostaining, Expressing, Cell Culture, Derivative Assay

Journal: Materials Today Bio

Article Title: Injectable deferoxamine-loaded microsphere hydrogels for inhibition of ferroptosis and promotion of third-degree burn wound healing

doi: 10.1016/j.mtbio.2025.101806

Figure Lengend Snippet: Inhibition of ferroptosis by DFO@GM-H hydrogels. (a) Representative fluorescent images of intracellular Fe 2+ accumulation in HSFs exposed to different groups for 24 h. Intracellular Fe 2+ were stained with FerroOrange in red fluorescence. (b) Quantitative analysis of intracellular Fe 2+ accumulation in HSFs. (c) Quantitative analysis of MDA content in HSFs exposed to different groups for 24 h. (d) Protein expression of ACSL4, SLC7A11, GPX4 and GAPDH in HSFs treated with different groups by western blot. (e–g) Quantification of fold-change of ACSL4, SLC7A11 and GPX4 in HSFs by western blot. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, ns means non-significant (p > 0.05).

Article Snippet: Tissue sections were incubated with primary antibodies overnight at 4 °C, including IL-1β mouse mAb (Proteintech, China, 66737-1-Ig, 1:300), Anti-CD31 rabbit mAb (Abcam, England, ab182981, 1:300), VEGFA rabbit pAb (Proteintech, China, 19003-1-ap, 1:500), ferritin light chain rabbit pAb ((Proteintech, China, 10727-1-AP, 1:500), ACSL4 rabbit mAb (Abcam, England, ab155282, 1:500), SLC7A11 rabbit mAb (Abcam, England, ab307601, 1:500), GPX4 mouse mAb (BOSTER, China, BA3802-1, 1:500).

Techniques: Inhibition, Staining, Fluorescence, Expressing, Western Blot

Journal: Materials Today Bio

Article Title: Injectable deferoxamine-loaded microsphere hydrogels for inhibition of ferroptosis and promotion of third-degree burn wound healing

doi: 10.1016/j.mtbio.2025.101806

Figure Lengend Snippet: Immunofluorescence staining and quantification of ferroptosis at burn wound sites. (a) Representative fluorescent images of intracellular Fe 2+ accumulation in skin tissues on day 8 post-wounding. (b) Representative fluorescent staining images of ACLS4, SLC7A11 and GPX4 in skin tissues on day 29 post-wounding. (c) Quantitative analysis of intracellular Fe 2+ level in skin tissues on day 8 post-wounding. (d–f) Quantitative analysis of ACLS4, SLC7A11 and GPX4 levels of skin tissues on days 29 post-wounding. ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, ns means non-significant (p > 0.05).

Article Snippet: Tissue sections were incubated with primary antibodies overnight at 4 °C, including IL-1β mouse mAb (Proteintech, China, 66737-1-Ig, 1:300), Anti-CD31 rabbit mAb (Abcam, England, ab182981, 1:300), VEGFA rabbit pAb (Proteintech, China, 19003-1-ap, 1:500), ferritin light chain rabbit pAb ((Proteintech, China, 10727-1-AP, 1:500), ACSL4 rabbit mAb (Abcam, England, ab155282, 1:500), SLC7A11 rabbit mAb (Abcam, England, ab307601, 1:500), GPX4 mouse mAb (BOSTER, China, BA3802-1, 1:500).

Techniques: Immunofluorescence, Staining