Journal: The Journal of Clinical Investigation

Article Title: Wilms tumor 1 impairs apoptotic clearance of fibroblasts in distal fibrotic lung lesions

doi: 10.1172/JCI188819

Figure Lengend Snippet: ( A ) Schematic representation of the animal experiment involving PDGFR α CreERT (control) and PDGFR α CreERT WT1 OE (cWT1 OE ) mice. Mice were treated repeatedly with bleomycin via intratracheal administration and with tamoxifen via intraperitoneal injection. ( B ) Quantification of Wt1 gene transcripts by RT-PCR in total lung RNA isolated from control and cWT1 OE mice ( n = 6/group). ** P < 0.01, by 2-tailed Student’s t test. ( C ) Representative confocal images of lung sections from control and cWT1 OE mice co-immunostained for WT1 (red), vimentin (green), and DAPI (blue). Scale bars: 20 μm. ( D ) Representative images of Masson’s trichrome–stained lung sections from control and cWT1 OE mice. Original magnification, ×4 and ×20, respectively. Scale bars: 1,500 μm and 200 μm, respectively. ( E ) The percentage of fibrotic area was quantified in control and cWT1 OE mice using BZ-X image analysis ( n = 6/group). **** P < 0.0001, by 2-tailed Student’s t test. ( F ) Hydroxyproline levels were measured in the right lungs of control and cWT1 OE mice ( n = 6/group). *** P < 0.001, by 2-tailed Student’s t test. ( G ) Lung resistance was assessed using FlexiVent in control and cWT1 OE mice treated with bleomycin ( n = 6/group). * P < 0.01, by 2-tailed Student’s t test. ( H ) Proliferation of fibroblasts was quantified using a BrdU incorporation assay in lung cultures from control and cWT1 OE mice treated with bleomycin ( n = 3/group). * P < 0.05, by 2-tailed Student’s t test. ( I ) Fibroblasts from the lung cultures of control and cWT1 OE mice treated with bleomycin were treated with anti-Fas antibody for 24 hours, followed by TUNEL staining (red). Representative confocal images are shown; nuclei are stained with DAPI (blue). Original magnification, ×20. Scale bars: 100 μm. ( J ) Percentage of TUNEL + fibroblasts in total DAPI + fibroblasts ( n = 3/group). * P < 0.05 and ** P < 0.01, by 1-way ANOVA. Data are representative of 2 independent experiments with similar findings.

Article Snippet: Cell proliferation was evaluated using the BrdU kit (Cell Signaling Technology) as described previously ( ).

Techniques: Control, Injection, Reverse Transcription Polymerase Chain Reaction, Isolation, Staining, BrdU Incorporation Assay, TUNEL Assay

Journal: The veterinary quarterly

Article Title: Multiple roles of LncRNA-BMNCR on cell proliferation and apoptosis by targeting miR-145/CBFB axis in BMECs.

doi: 10.1080/01652176.2023.2262525

Figure Lengend Snippet: Figure 2. BMNCR Facilitated proliferation and attenuated apoptosis in BMECs. (A) qRT-PCR detected the expression efficiency of BMNCR after transfected three BMNCR siRNAs, respectively. (B) The expression levels of inflammation-related cytokines were validated by RT‐qPCR after treated siBMNCR for 48 h. (C) CCK8 assay exploring the function of BMNCR on the viability of BMECs. (D) EdU assay detected the number of BMECs treated with siBMNCR. (E) The proportion of EdU positive cells was counted by ImageJ. (F) Cell apoptosis was determined by flow cytometry after transfected with siBMNCR. (G) Distribution map of BMECs apoptosis. (H) Cell apoptosis index was counted by the sum of early and late apoptosis. Data are means ± SE of n = 3 independent experiments, each performed in triplicate, and normalized to GAPDH. *, p < .05 and **, p < .01.

Article Snippet: Finally, cells were stained using the Cell Proliferation Assay Kit (C0075S, Beyotime, Shanghai, China), and images were collected on the Inverted Fluorescence Microscope (DMi8, Leica, Germany).

Techniques: Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, EdU Assay, Flow Cytometry

Journal: The veterinary quarterly

Article Title: Multiple roles of LncRNA-BMNCR on cell proliferation and apoptosis by targeting miR-145/CBFB axis in BMECs.

doi: 10.1080/01652176.2023.2262525

Figure Lengend Snippet: Figure 4. Functional roles of miR-145 on proliferation and apoptosis of BMECs. (A) qRT-PCR detected the overexpression (mimic) or Interference (inhibitor) efficiency of miR-145. The expression levels of inflammation-related cytokines were validated by RT‐qPCR after treated miR-145 mimic (B) or miR-145 inhibitor (C). (D) EdU assay detected the number of BMECs after treated with miR-145 mimic or inhibitor. (E) The proportion of EdU positive cells was counted by ImageJ. (F) Cell apoptosis was determined by flow cytometry after transfected with miR-145 mimic or inhibitor. (G) Distribution map of BMECs apoptosis. (H) Cell apoptosis index was counted by the sum of early and late apoptosis. Data are means ± SE of n = 3 independent exper iments, each performed in triplicate, and normalized to GAPDH. *, p < .05 and **, p < .01.

Article Snippet: Finally, cells were stained using the Cell Proliferation Assay Kit (C0075S, Beyotime, Shanghai, China), and images were collected on the Inverted Fluorescence Microscope (DMi8, Leica, Germany).

Techniques: Functional Assay, Quantitative RT-PCR, Over Expression, Expressing, EdU Assay, Flow Cytometry, Transfection

Journal: The veterinary quarterly

Article Title: Multiple roles of LncRNA-BMNCR on cell proliferation and apoptosis by targeting miR-145/CBFB axis in BMECs.

doi: 10.1080/01652176.2023.2262525

Figure Lengend Snippet: Figure 7. CBFB could modulate proliferation and apoptosis in BMECs. (A) qRT-PCR detected the expression efficiency of CBFB after transfected three siRNAs of CBFB, respectively. After transfected siCBFB into BMECs for 48 h, (B) mRNA level of CBFB was explored by qRT-PCR. (C) The protein level of CBFB was explored by Western blot. (D) The relative protein level of CBFB was calculated by ImageJ. (E) The expression levels of inflammation-related cytokines were validated by RT‐qPCR after treated siCBFB into BMECs for 48h. (F) EdU assay detected the number of BMECs treated with siCBFB. (G) The proportion of EdU positive cells was counted by ImageJ. (H) Cell apoptosis was determined by flow cytometry after transfected with siCBFB. (I) Distribution map of BMECs apoptosis. (J) Cell apoptosis index was counted by the sum of early and late apoptosis. Data are means ± SE of n = 3 independent experiments, each performed in triplicate, and normalized to GAPDH. *, p < .05 and **, p < .01.

Article Snippet: Finally, cells were stained using the Cell Proliferation Assay Kit (C0075S, Beyotime, Shanghai, China), and images were collected on the Inverted Fluorescence Microscope (DMi8, Leica, Germany).

Techniques: Quantitative RT-PCR, Expressing, Transfection, Western Blot, EdU Assay, Flow Cytometry