Journal: Cell reports

Article Title: Pituitary adenomas evade apoptosis via noxa deregulation in Cushing’s disease

doi: 10.1016/j.celrep.2022.111223

Figure Lengend Snippet: (A) A sublabial transsphenoidal approach was used for resection of microadenomas (white arrowheads) in patients with Cushing’s disease (CD). Coronal, post-gadolinium contrast enhanced magnetic resonance image from patient P1. Adenoma and adjacent normal pituitary gland were separately annotated during surgery and dissociated into a single-cell suspension, followed by GEM embedding, sequencing, and computational analysis. Scale bar: 2 cm. (B) Summary of demographic data and analysis for patients included in the study. (C) UMAP embedding of cells from the 6 patient samples with CD, GH, NFPA, and PRL adenomas used in scRNA-seq analysis. Cells cluster according to dominant secretory phenotype. (D) UMAP embedding showing the cell-type identities cells from the 6 patient samples. (E) UMAP embedding showing CD sample identity for cells from patients P1, P2, and P3. (F) UMAP plot showing CD patient cells clustering by cell-type identity. Cell types: leukocytes (Les), endothelial cells (ECs), pericytes (Pes), folliculostellate cells (FSs), corticotrophs (Cs), gonadotroph (Gs), somatotrophs (Ss), lactotrophs (Ls), ambiguous/somato-lactotrophs (SLs), and POU1F1 -positive, hormone-negative (P + H − ) cells. GH, growth hormone; NFPA, non-functioning pituitary adenoma; PRL, prolactinoma.

Article Snippet: GH1 , Novus Biologicals , Cat #NBP2–53262.

Techniques: Suspension, Sequencing

Journal: Cell reports

Article Title: Pituitary adenomas evade apoptosis via noxa deregulation in Cushing’s disease

doi: 10.1016/j.celrep.2022.111223

Figure Lengend Snippet: (A) The UMAP plot from split by patient (top, P1; middle, P2; bottom, P3), colored by cell type. (B) Dot plot showing the expression (marker color) and percentage of cells expressing (marker size) for the top 6 cell-type upregulated genes with highest min.logFC.detected from a filtered group of robustly upregulated cell-type marker genes . Genes used as a priori known classification marker genes are indicated by bold font. (C) Multiplexed immunohistochemistry of tissue from P3, showing localization of key hormone markers in margin and core regions of the sample. White bar: 100 μm. PC, pituitary cells; HPC, hormone-producing cells; Mar, tumor margin; POMC, pro-opiomelanocortin; GH, growth hormone; PRL, prolactin; LH, luteinizing hormone; FSH, follicle-stimulating hormone.

Article Snippet: GH1 , Novus Biologicals , Cat #NBP2–53262.

Techniques: Expressing, Marker, Immunohistochemistry

Journal: Cell reports

Article Title: Pituitary adenomas evade apoptosis via noxa deregulation in Cushing’s disease

doi: 10.1016/j.celrep.2022.111223

Figure Lengend Snippet: (A) UMAP plot demonstrating PMAIP1 abundance in CD adenomas (C) but not in PRL, G, or NFPA adenomas. PMAIP1 was also detected at lower levels in Les and ECs. (B) UMAP plot showing MYC upregulation in CD corticotrophs but not other hormone-producing cells (left panel). MYC expression was also detected in Pes, ECs, and Les. Middle panel: UMAP plot showing overlap of MYC and PMAIP1 expression is mostly limited to CD corticotrophs (yellow dots). Right panel: UMAP plot showing UCHL1 abundance in most hormone-producing cell types in CD. (C) Bulk RNA-seq of CD and non-CD samples verified overexpression of pro-apoptotic genes including PMAIP1 in CD tissues. (D) DNA methylation levels (beta values) at CpG sites associated with the PMAIP1 promoter methylation demonstrating hypomethylation in CD (n = 3) compared with normal (autopsy-derived, n = 20) pituitary glands. *p < 0.05. (E) Multiplex immunohistochemistry (mIHC) of 5 μm thick sections from a CD adenoma. Insets from the core-margin boundary represented by white dashed lines. White bar: 100 μm. Core-margin boundary identified by overlaying expression of POMC, TBX19, and DAPI. Compared with the margin, core adenoma cells show robust overexpression of POMC, TBX19, and MYC; however, noxa expression is decreased within the adenoma core. (F) Representative image from noxa IHC in independent adenoma/normal pairs (n = 10). Pairwise analysis of noxa deconvoluted IHC images (absorbance = mean pixel intensity count per pixel) showing suppressed noxa signal in CD adenomas compared with normal (margin) tissues (p = 0.0013; 95% confidence interval [CI] −0.027 to −0.007). Scale bar: 100 μM. (G) Expected epithelial growth factor (EGF) signaling upregulation and ERK1/2 phosphorylation were found in human CD adenoma primary cell lines (P6_CD and P26_CD). A, adenoma (core); N, normal (margin) pituitary gland. Noxa was undetectable or decreased in core adenomas. (H) A survey of human primary CD adenoma cell lines revealed variable noxa expression compared with sCD adenoma, GH adenoma, and NFPA. CD, corticotroph adenoma causing Cushing’s disease; sCD, hormonally silent corticotroph adenoma; GH, growth-hormone-secreting adenoma; NFPA, non-functioning pituitary adenoma.

Article Snippet: GH1 , Novus Biologicals , Cat #NBP2–53262.

Techniques: Expressing, RNA Sequencing, Over Expression, DNA Methylation Assay, Methylation, Derivative Assay, Multiplex Assay, Immunohistochemistry, Phospho-proteomics

Journal: Cell reports

Article Title: Pituitary adenomas evade apoptosis via noxa deregulation in Cushing’s disease

doi: 10.1016/j.celrep.2022.111223

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: GH1 , Novus Biologicals , Cat #NBP2–53262.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Sequencing

Journal: Cell stem cell

Article Title: Super-Obese Patient-Derived iPSC Hypothalamic Neurons Exhibit Obesogenic Signatures and Hormone Responses

doi: 10.1016/j.stem.2018.03.009

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: GhrR , Alomone Labs , Cat# AGR031 RRID: AB_2340976.

Techniques: Recombinant, Electron Microscopy, Avidin-Biotin Assay, Enzyme-linked Immunosorbent Assay, Protease Inhibitor, Microelectrode Array, Magnetic Beads, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Sequencing, Software

Journal: FEBS Open Bio

Article Title: Growth hormone receptor promotes osteosarcoma cell growth and metastases

doi: 10.1002/2211-5463.12761

Figure Lengend Snippet: GHR promotes OS cell colony formation and OS tumor growth through the PI3K/AKT pathway. (A) The expression of GHR was detected by western blotting. (B) Control and GHR knockdown 143B and U2OS cells were analyzed by colony formation. (C) Statistical analysis of the colony formation in (B). Data represent mean ± SEM ( n = 3); two‐tailed Student’s t ‐test was used for statistical analysis, ** P < 0.01, *** P < 0.001 in silenced GHR cells (D, E) Knockdown of GHR by two independent siRNAs inhibited the expression of p‐PI3K/AKT in 143B (D) and U2OS (E) cells.

Article Snippet: The used antibodies were as follows: actin (Sigma‐Aldrich, St. Louis, MO, USA), GHR (Proteintech, Shanghai, China), p‐PI3K (Cell Signaling, Shanghai, China), p‐AKT (Cell Signaling), PI3K (Cell Signaling), AKT (Cell Signaling), Goat anti‐Rabbit (Thermo, Danvers, MA, USA) and Goat anti‐Mouse (Thermo).

Techniques: Expressing, Western Blot, Control, Knockdown, Two Tailed Test

Journal: FEBS Open Bio

Article Title: Growth hormone receptor promotes osteosarcoma cell growth and metastases

doi: 10.1002/2211-5463.12761

Figure Lengend Snippet: GHR modulates the proliferation of OS cells in vivo . (A–C) Control and GHR knockdown U2OS cells were injected into nude mice subcutaneously (2 × 10 6 cells per mouse); tumor volume and weight were measured at the indicated day. Data represent mean ± SEM ( n = 5); two‐tailed Student’s t ‐test was used for statistical analysis, *** P < 0.001. (D) Western blotting analysis of p‐PI3K/AKT for GHR knockdown (sh GHR) or control (sh NC) tumors.

Article Snippet: The used antibodies were as follows: actin (Sigma‐Aldrich, St. Louis, MO, USA), GHR (Proteintech, Shanghai, China), p‐PI3K (Cell Signaling, Shanghai, China), p‐AKT (Cell Signaling), PI3K (Cell Signaling), AKT (Cell Signaling), Goat anti‐Rabbit (Thermo, Danvers, MA, USA) and Goat anti‐Mouse (Thermo).

Techniques: In Vivo, Control, Knockdown, Injection, Two Tailed Test, Western Blot

Journal: Molecular Medicine

Article Title: Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer

doi: 10.1186/s10020-025-01206-5

Figure Lengend Snippet: HM13 cleaves the signal peptide, resulting in changes in GGH localization and protein function. ( A ) The binding sites of the GGH gene sequence with signal peptidase were predicted using SignalP 6.0 prediction analysis. ( B ) GGH protein expression in HCT-15 cells. ( C ) GGH protein expression in HM13-knockdown HCT-15 cells. KD: knockdown. ( D ) Schematic diagram of GGH protein domain. ( E ) The total proteins, the proteins localized in lysosomes, the proteins localized in cytoplasm and nucleus were collected by Lysosome isolation experiment. Expression of GGH protein in the lysosomes and cytoplasm of IR group, IR + HM13 KD group and sham IR group. ( F ) Localization of GGH protein in IR group, IR + HM13 KD group and sham IR group. Red: lysosome; Green: GGH; Blue: DNA. ( G-I ) Levels of 5,10-CH 2 -THF polyglutamated metabolites (5,10-CH 2 -H 4 PteGlu n ) in various HCT-15 cells. Various types of 5,10-CH 2 -H 4 PteGlu n ( G ); Total 5,10-CH 2 -H 4 PteGlu ( H ); Proportion analysis of various types of 5,10-CH 2 -H 4 PteGlu ( I ). ( J-K ) Multiple gradient folic acid supplementation (maximum concentration 5 mM, 3×gradient dilution) after folic acid deprivation in the various tested groups. Folate growth requirement of IR group, IR + HM13 KD group and sham IR group, the dose–response curve: J ; the bar graph of IC 50 values: K . ( L-M ) Cell viability with 5-FU or 5-FU + 5,10-CH 2 -THF exposure in various HCT-15 cells, the dose–response curve: L ; the bar graph of IC 50 values: M . All figures were representatives of three independent experiments. Error bars represent the SD. **** P < 0.0001; ns: not significant, two-tailed Student’s t-test

Article Snippet: Primary antibodies used for western blot were: HM13 (ab190253, Abcam, UK), GGH (13264-1-AP, Proteintech, USA), GAPDH (60004-1-Ig, Proteintech, USA), ACTIN (66009-1-Ig, Proteintech, USA), LAMP-2 (SC-18822, Santa Cruz, USA), Tubulin (10068-1-AP, Proteintech, USA).

Techniques: Binding Assay, Sequencing, Expressing, Knockdown, Isolation, Concentration Assay, Two Tailed Test

Journal: Molecular Medicine

Article Title: Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer

doi: 10.1186/s10020-025-01206-5

Figure Lengend Snippet: 5-FU-resistance can be overcome by the combination of Met in vivo. ( A ) Scheme for 5-FU, Met, and combination therapy after irradiation treatment in the HCT-15-injection nude mouse. ( B-D ) Levels 5,10-CH 2 -H 4 PteGlu n in IR group and sham IR group HCT-15-injection nude mouse. Various types of 5,10-CH 2 -H 4 PteGlu n ( B ); Total 5,10-CH 2 -H 4 PteGlu ( C ); Proportion analysis of various types of 5,10-CH 2 -H 4 PteGlu ( D ). ( E ) Endogenous GGH and HM13 protein expression in IR group, IR + HM13 KD group and sham IR group HCT-15-injection nude mouse. ( n = 4). ( F-H ) Growth curve ( F ) and photographs ( G ) of tumors collected from mice treated with 5-FU, Met or 5-FU + Met; Tumor weight at the endpoint of the in vivo experiment ( H ). ( I-K ) Growth curve ( I ) and photographs ( J ) of tumors collected from mice-irradiated (2 Gy × 8) treated with 5-FU, Met and the combination; Tumor weight at the endpoint of the in vivo experiment ( K ). ( n = 4). ( L ) Effect of 5-FU and Met combined therapy (post-IR Day 24). Error bars represent the SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, two-tailed Student’s t-test

Article Snippet: Primary antibodies used for western blot were: HM13 (ab190253, Abcam, UK), GGH (13264-1-AP, Proteintech, USA), GAPDH (60004-1-Ig, Proteintech, USA), ACTIN (66009-1-Ig, Proteintech, USA), LAMP-2 (SC-18822, Santa Cruz, USA), Tubulin (10068-1-AP, Proteintech, USA).

Techniques: In Vivo, Irradiation, Injection, Expressing, Two Tailed Test

Journal: Human Vaccines & Immunotherapeutics

Article Title: Cross-reactive humoral immunity of clade 2 Oka and MAV/06 strain-based varicella vaccines against different clades of varicella–zoster virus

doi: 10.1080/21645515.2023.2210961

Figure Lengend Snippet: Image of FAMA test with anti-VZV glycoprotein monoclonal antibodies using six different VZV strains as FAMA antigens (400× magnification). (a) gH, (b) gB, (c) gI, (d) gE.

Article Snippet: Sera were twofold serially diluted with Dulbecco’s phosphate-buffered saline (DPBS, Lonza, Breda, Netherlands), and mouse monoclonal anti-VZV glycoprotein H (gH), gB, gI, and gE antibodies (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) were diluted 1:50 in DPBS.

Techniques: Bioprocessing