normal human osteoblasts  (Lonza)

Journal: Non-coding RNA Research

Article Title: Long noncoding RNA LINC00339 promotes osteoporosis development via modulating of regulator CDC42 by binding PARP1

doi: 10.1016/j.ncrna.2025.06.004

Figure Lengend Snippet: Osteoblast function is closely correlated with LINC00339 expression. A. The expression of total LINC00339 in the bone tissue of healthy controls (n = 15) and patients with osteoporosis (n = 18). B. Correlation analysis between LINC00339 and T score in bone specimens. C. Analysis of the expression of the osteogenic marker genes OCN after LINC00339 knockdown in primary human osteoblasts. D, E. Analysis of the expression of the osteogenic marker genes RUNX2, ALP and OCN after LINC00339 knockdown and overexpression in U2OS cells. Representative results of three independent experiments are shown. F. ALP staining images and representative Alizarin red staining images of the sh-NC and sh-LINC00339 osteoblasts induced with osteogenic medium for 7 days. Scale bar, 5 mm. G . Quantification of ALP staining areas (Left) and ARS staining areas (Right). H . ALP staining images and representative ARS images of the OE-NC and OE-LINC00339 osteoblasts induced with osteogenic medium for 7 days. Scale bar, 5 mm. I . Quantification of ALP staining areas (Left) and ARS staining areas (Right). n = 3 for each group. Data are represented as mean ± standard deviation. Two-way ANOVA was performed to study the interaction between two independent variables. Significances were determined using two-tailed paired student's t -test between two groups. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: The lentiviruses generated and released into the supernatant of the cultured HEK293T cells, these were subsequently collected and concentrated, then add condensed lentiviruses into U2OS and human primary osteoblasts cells in the presence of polybrene (Solarbio, China).

Techniques: Expressing, Marker, Knockdown, Over Expression, Staining, Standard Deviation, Two Tailed Test

Journal: Non-coding RNA Research

Article Title: Long noncoding RNA LINC00339 promotes osteoporosis development via modulating of regulator CDC42 by binding PARP1

doi: 10.1016/j.ncrna.2025.06.004

Figure Lengend Snippet: LINC00339 functions by interacting with PARP1. A. RNA FISH showed that LINC00339 was predominantly localized in the cytoplasm. Scale bar, 50 μm. B. Schematic diagram of RNA pull-down experiment. The antisense and sense of LIN00339 were synthesized in vitro. C. Silver staining of biotinylated LIN00339-associated proteins. LC-MS identified the differential RBPs binding to AS-LINC00339 and S-LINC00339. D. Western blot of protein from LINC00339-pulldown assays. Western blot with PARP1 antibody shows only the sense of LINC00339 enrichment PARP1. E. PARP1 RIP assay to analyze interactions between PARP1 and LINC00339 in U2OS osteoblast-like cells. WB shows the PARP1 antibody efficiency of immunoprecipitation. F. RT-qPCR to analyze the enrichment of LINC00339 in RNA-protein complexes. The LINC00339 abundance in anti-PARP1 group was much more than the IgG group. Values were normalized by the input group. ∗∗∗ P < 0.001 versus IgG, by Student's t -test. (AS-LINC00339: Anti-sense of LINC00339, S-LINC00339: Sense LINC00339). G . IF-RNA FISH experiment was performed in U2OS cells to detect the co-localization of LINC00339 and PARP1. Bar: 10 μm. Data are expressed as the mean ± standard deviation of 3 independent experiments. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001, two-tailed paired student's t -test.

Article Snippet: The lentiviruses generated and released into the supernatant of the cultured HEK293T cells, these were subsequently collected and concentrated, then add condensed lentiviruses into U2OS and human primary osteoblasts cells in the presence of polybrene (Solarbio, China).

Techniques: Synthesized, In Vitro, Silver Staining, Liquid Chromatography with Mass Spectroscopy, Binding Assay, Western Blot, Immunoprecipitation, Quantitative RT-PCR, Standard Deviation, Two Tailed Test

Journal: Non-coding RNA Research

Article Title: Long noncoding RNA LINC00339 promotes osteoporosis development via modulating of regulator CDC42 by binding PARP1

doi: 10.1016/j.ncrna.2025.06.004

Figure Lengend Snippet: LINC00339–PARP1 complex co-regulates the expression of CDC42. A. The effects of LINC00339 knockdown on PARP1 mRNA expression were detected in primary human osteoblasts. B. Effect of LINC00339 overexpression on mRNA and protein expression of PARP1 in U2OS cells. C. Effect of PARP1 knockdown on LINC00339 overexpression-induced mRNA expression of CDC42. D. Effect of PARP1 knockdown on LINC00339 overexpression-induced protein expression of CDC42. E, F. Effect of PARP1 overexpression on LINC00339 knockdown-induced mRNA and protein expressions of CDC42. Data are expressed as the mean ± standard deviation of 3 independent experiments. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 versus vector, by Student's t -test.

Article Snippet: The lentiviruses generated and released into the supernatant of the cultured HEK293T cells, these were subsequently collected and concentrated, then add condensed lentiviruses into U2OS and human primary osteoblasts cells in the presence of polybrene (Solarbio, China).

Techniques: Expressing, Knockdown, Over Expression, Standard Deviation, Plasmid Preparation

Journal: Non-coding RNA Research

Article Title: Long noncoding RNA LINC00339 promotes osteoporosis development via modulating of regulator CDC42 by binding PARP1

doi: 10.1016/j.ncrna.2025.06.004

Figure Lengend Snippet: Proposed model for LINC00339-mediated regulation of the differentiation in osteoblast.

Article Snippet: The lentiviruses generated and released into the supernatant of the cultured HEK293T cells, these were subsequently collected and concentrated, then add condensed lentiviruses into U2OS and human primary osteoblasts cells in the presence of polybrene (Solarbio, China).

Techniques: