control scrambled shrna sh nc  (Shanghai Genechem Ltd)

Journal: Aging Cell

Article Title: BRD4 / MAP2K7 / PGF Signaling Axis Promotes Senescence and Extracellular Matrix Metabolism of Nucleus Pulposus Cells in Intervertebral Disk Degeneration

doi: 10.1111/acel.70034

Figure Lengend Snippet: BRD4 mediates NP cell senescence and ECM metabolism. (a) Western blot confirming the knockdown of BRD4 in NP cells post‐transfection with BRD4 shRNA lentivirus. (b) Western blot showing BRD4 overexpression in NP cells following transfection with BRD4 gene‐sequence lentivirus (LV‐BRD4). (c, d) Representative images and quantitative analysis of SA‐β‐gal staining to evaluate the extent of senescence across different groups. The intensity of blue staining serves as an indicator of elevated SA‐β‐gal activity, suggesting a higher degree of cellular senescence. (e–h) Representative Western blot images and quantitative analysis of P16, P21, IL‐6, IL‐8, MMP3, MMP13, collagen II, and aggrecan expression levels in NP cells following BRD4 knockdown and overexpression. (i, j) Representative images and quantitative analysis of immunofluorescence staining for BRD4, P16, aggrecan, and MMP3 across different groups. (k, l) Representative images and quantitative analysis of Safranin O staining in various groups. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Lentiviruses expressing short hairpin RNA (shRNA) targeting BRD4 (sh‐BRD4), MAP2K7 (sh‐MAP2K7), and PGF (sh‐PGF), along with scrambled control shRNA (sh‐NC) and lentiviruses overexpressing BRD4 (LV‐BRD4), MAP2K7 (LV‐MAP2K7), PGF (LV‐PGF), and a control lentivirus (LV‐NC) were purchased from GeneChem (Shanghai, China).

Techniques: Western Blot, Knockdown, Transfection, shRNA, Over Expression, Sequencing, Staining, Activity Assay, Expressing, Immunofluorescence

Journal: Aging Cell

Article Title: BRD4 / MAP2K7 / PGF Signaling Axis Promotes Senescence and Extracellular Matrix Metabolism of Nucleus Pulposus Cells in Intervertebral Disk Degeneration

doi: 10.1111/acel.70034

Figure Lengend Snippet: MAP2K7 mediates NP cell senescence and ECM metabolism. (a, b) Representative immunohistochemical images and quantitative analysis of MAP2K7 expression in NP tissues from Grade II and Grade V patients with IDD, as well as from young (2 M) and aged (20 M) rats. (c, d) Representative Western blot bands and quantitative analysis of MAP2K7 expression in NP tissues from Grade II and Grade V patients, and from young (2 M) and aged (20 M) rats. (e) Western blot validation showing MAP2K7 knockdown in NP cells after transfection with MAP2K7 shRNA lentivirus and overexpression after transfection with MAP2K7 gene‐sequence lentivirus (LV‐MAP2K7). (f, g) Representative images and quantitative analysis of SA‐β‐gal staining used to evaluate the level of senescence across different intervention groups. Increased blue staining reflects higher SA‐β‐gal activity, indicating greater cellular senescence. (h–k) Representative Western blot bands and quantitative analysis of P16, P21, IL‐6, IL‐8, MMP3, collagen II, and aggrecan expression levels following MAP2K7 knockdown and overexpression in NP cells. (l, m) Representative images and quantitative analysis of immunofluorescence staining for MAP2K7, P16, aggrecan, and MMP3 in NP cells from different groups. (n, o) Representative images and quantitative analysis of Safranin O staining in different groups. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Lentiviruses expressing short hairpin RNA (shRNA) targeting BRD4 (sh‐BRD4), MAP2K7 (sh‐MAP2K7), and PGF (sh‐PGF), along with scrambled control shRNA (sh‐NC) and lentiviruses overexpressing BRD4 (LV‐BRD4), MAP2K7 (LV‐MAP2K7), PGF (LV‐PGF), and a control lentivirus (LV‐NC) were purchased from GeneChem (Shanghai, China).

Techniques: Immunohistochemical staining, Expressing, Western Blot, Biomarker Discovery, Knockdown, Transfection, shRNA, Over Expression, Sequencing, Staining, Activity Assay, Immunofluorescence

Journal: Aging Cell

Article Title: BRD4 / MAP2K7 / PGF Signaling Axis Promotes Senescence and Extracellular Matrix Metabolism of Nucleus Pulposus Cells in Intervertebral Disk Degeneration

doi: 10.1111/acel.70034

Figure Lengend Snippet: PGF mediates NP cell senescence and ECM metabolism. (a, b) Representative immunohistochemical images and quantitative analysis of PGF expression in NP tissues from young (2 months, 2 M) and aged (20 months, 20 M) rats. (c) Quantitative analysis of immunohistochemical expression levels of PGF, BRD4, and MAP2K7. (d, e) Representative Western blot bands and quantitative analysis of PGF expression in NP tissues from young (2 M) and aged (20 M) rats. (f) Western blot validation of PGF knockdown in NP cells following transfection with PGF shRNA lentivirus, and overexpression after transfection with PGF gene‐sequence lentivirus (LV‐PGF). (g, h) Representative images and quantitative analysis of SA‐β‐gal staining to evaluate the level of senescence in different groups. The intensity of blue staining reflects higher SA‐β‐gal activity, indicating more pronounced cellular senescence. (i–l) Representative Western blot bands and quantitative analysis of P16, P21, IL‐6, IL‐8, collagen II, and aggrecan expression levels after PGF knockdown and overexpression in NP cells. (m, n) Representative images and quantitative analysis of immunofluorescence staining for P16, aggrecan, and MMP3 across different groups. (o, p) Representative images and quantitative analysis of Safranin O staining in different groups. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Lentiviruses expressing short hairpin RNA (shRNA) targeting BRD4 (sh‐BRD4), MAP2K7 (sh‐MAP2K7), and PGF (sh‐PGF), along with scrambled control shRNA (sh‐NC) and lentiviruses overexpressing BRD4 (LV‐BRD4), MAP2K7 (LV‐MAP2K7), PGF (LV‐PGF), and a control lentivirus (LV‐NC) were purchased from GeneChem (Shanghai, China).

Techniques: Immunohistochemical staining, Expressing, Western Blot, Biomarker Discovery, Knockdown, Transfection, shRNA, Over Expression, Sequencing, Staining, Activity Assay, Immunofluorescence