sirnas on-target and non-specific control  (Ribobio co)

Journal: Cell Death & Disease

Article Title: Fbxo22 promotes cervical cancer progression via targeting p57 Kip2 for ubiquitination and degradation

doi: 10.1038/s41419-022-05248-z

Figure Lengend Snippet: A , B FBXO22 knockdown increased p57 protein expression ( A ) and its ectopic expression reduced p57Kip2 protein expression ( B ) in SiHa and C33A cells detected by Western blotting analysis. Relative FBXO22 and p57Kip2 protein level is presented as normalized band density to the corresponding GAPDH band density. Each data in the plots is the mean ± SEM from three independent Western blots. * P < 0.05, ** P < 0.01. shCtr: control shRNA, shR2: shFBXO22-2, EV: empty vector, OE: FBXO22 overexpression. C , D FBXO22 knockdown ( C ) or overexpression ( D ) showed no effect on the p57Kip2 mRNA expression in SiHa and C33A cells. Each data in the plots are expressed as the means ± SEM from triplicate measurements. E , F FBXO22 and p57Kip2 proteins physically interact with each other in SiHa and C33A cells by IP assays as detected by immunoblotting (IB) for FBXO22 in the precipitated proteins by an anti-p57Kip2 antibody ( E ) and for p57Kip2 in the precipitated proteins by an anti-FBXO22 antibody ( F ).

Article Snippet: A pool of four small interfering RNAs (siRNAs) targeting p57 Kip2 and non-specific control (Ctr siRNA) were purchased from Thermo Fisher (Waltham, MA, USA) and transfected into the CC cells in the presence of Dharma (Thermo Fisher, USA).

Techniques: Knockdown, Expressing, Western Blot, Control, shRNA, Plasmid Preparation, Over Expression

Journal: Cell Death & Disease

Article Title: Fbxo22 promotes cervical cancer progression via targeting p57 Kip2 for ubiquitination and degradation

doi: 10.1038/s41419-022-05248-z

Figure Lengend Snippet: A , B SiHa cells with FBXO22 knockdown ( A ) or overexpression ( B ) were treated with cycloheximide (CHX) at 100 μM, and then harvested at the indicated time points. The cell lysate was analyzed by the Western blotting for FBXO22, p57Kip2, and GAPDH levels (left panel). Quantification of the p57 levels relative to GAPDH expression is also shown (right panel). C Western blotting was used to detect the changes of p57Kip2 protein in SiHa and C33A cells without or with FBXO22 knockdown before and after MG132 (20 μM) treatment. D The levels of ubiquitinated p57Kip2 was measured in immunoprecipitated p57Kip2 in the CC cells with FBXO22 knockdown or overexpression.

Article Snippet: A pool of four small interfering RNAs (siRNAs) targeting p57 Kip2 and non-specific control (Ctr siRNA) were purchased from Thermo Fisher (Waltham, MA, USA) and transfected into the CC cells in the presence of Dharma (Thermo Fisher, USA).

Techniques: Knockdown, Over Expression, Western Blot, Expressing, Immunoprecipitation

Journal: Cell Death & Disease

Article Title: Fbxo22 promotes cervical cancer progression via targeting p57 Kip2 for ubiquitination and degradation

doi: 10.1038/s41419-022-05248-z

Figure Lengend Snippet: A The protein levels of FBXO22 and p57Kip2 were detected by Western blotting in SiHa and C33A control (shCtr) and FBXO22 knockdown (shR2) cells transfected with a control siRNA (CtrsiRNA) or a p57Kip2 siRNA (p57siRNA). B , C MTT ( B ) and colony formation ( C ) assays were used to measure the viability and proliferation in SiHa and C33A cells with or without FBXO22 and/or p57 knockdown. D , E Cell cycle was analyzed in SiHa and C33A cells with or without FBXO22 and/or p57 knockdown. F Cell apoptosis was analyzed in SiHa and C33A cells with or without FBXO22 and/or p57 knockdown. Data are shown as mean ± SEM from triplicate measurements. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: A pool of four small interfering RNAs (siRNAs) targeting p57 Kip2 and non-specific control (Ctr siRNA) were purchased from Thermo Fisher (Waltham, MA, USA) and transfected into the CC cells in the presence of Dharma (Thermo Fisher, USA).

Techniques: Western Blot, Control, Knockdown, Transfection

Journal: Cell Death & Disease

Article Title: Fbxo22 promotes cervical cancer progression via targeting p57 Kip2 for ubiquitination and degradation

doi: 10.1038/s41419-022-05248-z

Figure Lengend Snippet: A Representative images of IHC staining with anti-FBXO22 and anti-p57Kip2 antibodies in tissue sections of the tumors formed by the control, FBXO22 knockdown (shR2), or overexpressing (OE) SiHa cells (×400, magnification). B Linear correlation analysis of FBXO22 and p57Kip2 IHC scores in mouse xenograft tissues. C Western blotting analysis of the FBXO22 and p57Kip2 protein levels in the tumors formed by the control, FBXO22 knockdown (shR2), or overexpressing (OE) SiHa cells. The relative FBXO22 and p57Kip2 protein level is presented as normalized band density to the corresponding GAPDH band density. Each data in the plots is the mean ± SEM from four randomly selected tumors. * P < 0.05, ** P < 0.01. D Linear correlation analysis of the relative FBXO22 and p57Kip2 protein levels in the selected mouse tumors formed by the control, FBXO22 knockdown (shR2), or overexpressing (OE) SiHa cells. E Representative images of Western blotting analysis of the FBXO22 and p57Kip2 protein levels in tumor-adjacent cervical tissues (N) and cervical cancer tissues (T) of 6 CC patients. F Relative expression of FBXO22 and p57Kip2 protein as normalized band density to the GAPDH band density. Each data in the plot is the mean ± SEM from 10 normal or tumor tissue samples. * P < 0.05. G Linear correlation analysis of the relative FBXO22 and p57 protein levels in 20 tissue samples.

Article Snippet: A pool of four small interfering RNAs (siRNAs) targeting p57 Kip2 and non-specific control (Ctr siRNA) were purchased from Thermo Fisher (Waltham, MA, USA) and transfected into the CC cells in the presence of Dharma (Thermo Fisher, USA).

Techniques: Immunohistochemistry, Control, Knockdown, Western Blot, Expressing