Journal: Allergy

Article Title: Human Pulmonary Neuroendocrine Cells Respond to House Dust Mite Extract With PAR-1 Dependent Release of CGRP.

doi: 10.1111/all.16416

Figure Lengend Snippet: FIGURE 1 | Characterization of iPNEC and ePNEC cultures. (A) Quantitative RT-PCR showing expression of characteristic PNEC markers in human cultured iPSC and -HBEC-derived iPNEC (□ female, 22; ■ male, 32) and ePNEC (● male, 52; ▲ male, 56; ○ female, 55) and control HBEC at day 60 in ALI culture, respectively. (B) Mean fluorescent intensity (MFI) of CHGA+/Hoechst+ cells (left) and SYP+/Hoechst+ cells [34] at day 60. Data is representative of four different culture plate wells per group for one biological donor for each cell type (ePNEC male, 52; iPNEC male, 32). (C) Representative IF images show SYP+ cells and nuclei are counterstained with Hoechst. 20× magnification and scale bars at 100 μm. (D) Single- cell RNA sequencing identifies two ePNEC clusters in 60-day-old ePNEC differentiated cells. Each dot represents one well and data shown for mean ± SEM. Mann–Whitney test performed for (A, B); * < 0.05. ASCL1, Achaete-Scute Family BHLH Transcription Factor 1; CHGA, Chromogranin A; ePNEC, epithelial-derived pulmonary neuroendocrine cells; iPNEC, iPSC-derived pulmonary neuroendocrine cells; iPSC, induced pluripotent stem cells; SYP, Synaptophysin.

Article Snippet: Cells were grown using the bronchial epithelial cell growth kit from ATCC (PCS300- 040).

Techniques: Quantitative RT-PCR, Expressing, Cell Culture, Derivative Assay, Control, RNA Sequencing, MANN-WHITNEY

Journal: Allergy

Article Title: Human Pulmonary Neuroendocrine Cells Respond to House Dust Mite Extract With PAR-1 Dependent Release of CGRP.

doi: 10.1111/all.16416

Figure Lengend Snippet: FIGURE 4 | CGRP release after HDM stimulation is PAR1-dependent (A) Single-cell transcriptomics data of iPNECs (GSE146990, Hor et al.) and ePNEC for PAR1 (F2R), PAR2 (F2RL1), PAR3 (F2RL2), and PAR4 (F2RL3) genes. (B) SYP and PAR1 immunohistochemical co-staining in naive iPNEC (male, 32). 20× and 100× magnification and scale bars at 50 and 10 μm, respectively. (C, D) CALCB mRNA (C) and CGRP protein (D) expression in different ePNEC (● male, 52; ▲ male, 56; ○ female, 55) conditions after 2 h: 50 μL PBS, HDM (1200 μg/mL), PAR1 inhibitor Vorapaxar (80 μM) and PAR1 agonist TFLLR-NH2 (2 μM). (E) CALCB mRNA RT-PCR expression in different ePNEC conditions after 2 h: 50 μL PBS, HDM (1200 μg/mL), HDM co-incubation with protease inhibitors Chymostatin (broad spectrum, 10 μg/mL), PMSF (serine specific, 0.25 mM) and E-64 (cysteine specific, 10 μM). Inhibitor/agonist and or HDM added at the same time. Each dot represents one well and data shown for mean ± SEM. One-way ANOVA for (C, D, E); * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. Symbol on top of multiple brackets represents all brackets. Agonist, PAR1 agonist vorapaxar; ePNEC, epithelial-derived pulmonary neuroendocrine cells; Inh., PAR1 inhibitor; iPNEC, iPSC-derived pulmonary neuroendocrine cells; HDM, House dust mite; PAR1, Protease activated receptor 1; SYP, Synaptophysin.

Article Snippet: Cells were grown using the bronchial epithelial cell growth kit from ATCC (PCS300- 040).

Techniques: Single-cell Transcriptomics, Immunohistochemical staining, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Incubation, Derivative Assay

Journal: Molecular medicine reports

Article Title: Intraperitoneal injection of thalidomide alleviates early osteoarthritis development by suppressing vascular endothelial growth factor expression in mice.

doi: 10.3892/mmr.2018.8980

Figure Lengend Snippet: Figure 2. mRNA expression levels of VEGF and MMP‑13 in the knee articular cartilage of mice among the Sham, Dmm and Dmm+Th groups (n=4 in each group). (A) Relative mRNA expression levels of VEGF in the medial articular cartilage. (B) Relative mRNA expression levels of MMP‑13 in the medial articular cartilage. The values are presented as the mean ± standard deviation. *P<0.05 compared with the Sham group; #P<0.05 compared with the Dmm group. Dmm, destabilization of the medial meniscus; MMP‑13, matrix metalloproteinase‑13; Th, thalidomide; VEGF, vascular endothelial growth factor.

Article Snippet: An ELISA kit of VEGF (E-EL-M1292c) was purchased from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Expressing, Standard Deviation

Journal: Molecular medicine reports

Article Title: Intraperitoneal injection of thalidomide alleviates early osteoarthritis development by suppressing vascular endothelial growth factor expression in mice.

doi: 10.3892/mmr.2018.8980

Figure Lengend Snippet: Figure 3. Immunohistochemical analysis of VEGF expression in the knee articular cartilage of mice among the Sham, Dmm and Dmm+Th groups (n=4 in each group). (A) Immunohistochemistry staining of VEGF in the articular cartilage of the medial tibial plateau (magnification, x400, scale bar=100 µm). (B) Quantification of VEGF positive cells, based on the results of immunohistochemistry staining. The values are presented as the mean ± standard deviation. *P<0.05 compared with the Sham group; #P<0.05 compared with the Dmm group. Dmm, destabilization of the medial meniscus; Th, thalidomide; VEGF, vascular endothelial growth factor.

Article Snippet: An ELISA kit of VEGF (E-EL-M1292c) was purchased from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Immunohistochemical staining, Expressing, Immunohistochemistry, Staining, Standard Deviation

Journal: Molecular medicine reports

Article Title: Intraperitoneal injection of thalidomide alleviates early osteoarthritis development by suppressing vascular endothelial growth factor expression in mice.

doi: 10.3892/mmr.2018.8980

Figure Lengend Snippet: Figure 5. ELISA analysis of serum VEGF concentration of mice among the Sham, Dmm and Dmm+Th groups (n=8 in each group). The values are presented as the mean ± standard deviation. *P<0.05 compared with the Sham group; #P<0.05 compared with the Dmm group. Dmm, destabilization of the medial meniscus; Th, thalidomide; VEGF, vascular endothelial growth factor.

Article Snippet: An ELISA kit of VEGF (E-EL-M1292c) was purchased from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Standard Deviation

Journal: Nature communications

Article Title: Mid-old cells are a potential target for anti-aging interventions in the elderly.

doi: 10.1038/s41467-023-43491-w

Figure Lengend Snippet: Fig. 2 | Mid-old specific gene expression in old-aged tissues. a–d Representative images of p21Waf1, SAA1, IL1β and SLIT2 IHC analysis in young and old-aged human colon and lung tissues are shown. “1” and “2” indicate high-magnification views of the original figure. Image “1” primarily depicts the epithelium and smooth muscle region, while image “2” depicts a stromal-rich region. Black arrows indicate stained cells in the stromal region. Yellow arrow in (d) indicate SLIT2-positive cells in

Article Snippet: Primary pulmonary artery smooth muscle cells (PASMCs) were purchased from American Tissue Culture Collections (ATCC, Rockville, MD) and were cultured in vascular cell basal medium (PCS-100-030, ATCC) supplemented with vascular smooth muscle cell growth kit (PCS-100-042, ATCC) at 37 °C in a humidified incubator with 5% CO2.

Techniques: Gene Expression, Staining