transwell inserts uncoated with matrigel  (Corning Life Sciences)

Journal: Cancer Cell International

Article Title: LINC00662 enhances cell progression and stemness in breast cancer by MiR-144-3p/ SOX2 axis

doi: 10.1186/s12935-022-02576-0

Figure Lengend Snippet: LINC00662 regulates BC cell progression by competitively binding to miR-144-3p. A Cytoplasmic and nuclear fraction RNA analysis was applied to detect LINC00662 location. B Putative miRNAs which share binding sites with LINC00662 were screened from starBase V2.0. C QRT-PCR analysis exhibited relative miRNAs expression after knockdown of LINC00662 . D RIP assay examined relative RNA expression. E Binding sites between LINC00662 wild type ( LINC00662 WT) and miR-144-3p and the base sequence of LINC00662 mutant type ( LINC00662 Mut) were demonstrated. Luciferase reporter assay verified the putative binding sites between LINC00662 and miR-144-3p. F CCK-8 assay was carried out to evaluate how cell proliferative ability changed after knockdown of LINC00662 and knockdown of LINC00662 and miR-144-3p both. G – I Transwell, sphere formation, and western blot assays were conducted to examine that inhibited cell migration, invasion, stemness ability and stemness-related protein level by knockdown of LINC00662 were rescued by knockdown of miR-144-3p. * p < 0.05; ** p < 0.01

Article Snippet: Matrigel-uncoated or Matrigel pre-coated transwell inserts (Millipore, Bedford, MA, USA) were employed for evaluation of cell migration or invasion.

Techniques: Binding Assay, Quantitative RT-PCR, Expressing, Knockdown, RNA Expression, Sequencing, Mutagenesis, Luciferase, Reporter Assay, CCK-8 Assay, Western Blot, Migration

Journal: Cancer Cell International

Article Title: LINC00662 enhances cell progression and stemness in breast cancer by MiR-144-3p/ SOX2 axis

doi: 10.1186/s12935-022-02576-0

Figure Lengend Snippet: LINC00662 targets miR-144-3p/ SOX2 axis to modulate BC cell progression. A QRT-PCR analysis was employed to investigate the expression of stemness-related genes in MCF-10A, MDA-MB-231, MCF-7, MDA-MB-468 and MDA-MB-453 cells. B Binding sites between SOX2 wild type ( SOX2 WT) and miR-144-3p and the base sequence of SOX2 mutant type ( SOX2 Mut) were demonstrated. C CCK-8 assay evaluated cell proliferative ability after the down-regulation of LINC00662 or knockdown of LINC00662 and overexpression of SOX2 . D – F Transwell, sphere formation, and western blot assays were conducted to examine cell migration, invasion, stemness ability and stemness-related protein level after the down-regulation of LINC00662 or knockdown of LINC00662 and overexpression of SOX2 . G RIP assay evaluated the enrichments of LINC00662 , miR-144-3p, SOX2 in RISC in BC cells. H Luciferase reporter assay was carried out to verify the ceRNA network among LINC00662 , miR-144-3p and SOX2 . ** p < 0.01; *** p < 0.001

Article Snippet: Matrigel-uncoated or Matrigel pre-coated transwell inserts (Millipore, Bedford, MA, USA) were employed for evaluation of cell migration or invasion.

Techniques: Quantitative RT-PCR, Expressing, Binding Assay, Sequencing, Mutagenesis, CCK-8 Assay, Knockdown, Over Expression, Western Blot, Migration, Luciferase, Reporter Assay

Journal: Cancer Cell International

Article Title: LINC00662 enhances cell progression and stemness in breast cancer by MiR-144-3p/ SOX2 axis

doi: 10.1186/s12935-022-02576-0

Figure Lengend Snippet: LINC00662 is overexpressed in BC tissues and promotes cell proliferation, migration, invasion and stemness. A QRT-PCR analysis was applied to detect LINC00662 expression in normal tissues and tumorous tissues. The mean, SD and 95% CI have been provided in Additional file : Table S4. B QRT-PCR analysis was applied to detect LINC00662 expression in MCF-10A, MDA-MB-231, MCF-7, MDA-MB-468 and MDA-MB-453. C QRT-PCR was employed to examine the efficiency of sh- LINC00662 #1/2/3. D Cell colony formation assay disclosed cell proliferative ability after the silencing of LINC00662 in MDA-MB-231 and MCF-7 cells. E Transwell assays were conducted to investigate cell migratory and invasive ability after knockdown of LINC00662 in BC cells. F Sphere formation assay was applied to detect cell stemness after LINC00662 knockdown in BC cells. G Western blot assay was conducted to examine the levels of stemness-related proteins after LINC00662 ablation in BC cells. ** p < 0.01

Article Snippet: Matrigel-uncoated or Matrigel pre-coated transwell inserts (Millipore, Bedford, MA, USA) were employed for evaluation of cell migration or invasion.

Techniques: Migration, Quantitative RT-PCR, Expressing, Colony Assay, Knockdown, Tube Formation Assay, Western Blot