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Association of CALB2 expression with <t>p53</t> signaling in colorectal adenocarcinoma. ( A , B ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( A ) and corresponding densitometric quantification ( B ) in HCT116 cells following CALB2 knockdown using two independent siRNAs (si1-CALB2 and si2-CALB2) or negative control (NC). ( C , D ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( C ) and corresponding densitometric quantification ( D ) in HT29 cells after CALB2 knockdown or negative control (NC). ( E , F ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( E ) and corresponding densitometric quantification ( F ) in SW48 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( G , H ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( G ) and corresponding densitometric quantification ( H ) in SW480 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( I – K ) RT-qPCR analysis of CALB2 ( I ), p21(CDKN1A) ( J ), and BAX ( K ) mRNA levels in HCT116 cells transfected with NC, si1-CALB2, or si1-CALB2 combined with the p53 inhibitor PFT-α (10 μM). ( L ) CCK-8 assay of HCT116 cells in the NC, si1-CALB2, and si1-CALB2+PFT-α groups over a 96 h period. ( M , N ) Representative images of scratch wound-healing assays ( M ) and quantitative analysis of relative migration rates ( N ) in HCT116 cells at 0, 24, and 48 h post-scratching. Scale bar = 250 μm. ( O – Q ) Representative double immunofluorescence staining ( O ) and quantitative analysis of fluorescence intensity ( P , Q ) for CALB2 and p-p53-Ser33 in xenograft tumor tissues from NC and OE-CALB2 groups. Scale bar = 50 μm. (n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).
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Association of CALB2 expression with <t>p53</t> signaling in colorectal adenocarcinoma. ( A , B ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( A ) and corresponding densitometric quantification ( B ) in HCT116 cells following CALB2 knockdown using two independent siRNAs (si1-CALB2 and si2-CALB2) or negative control (NC). ( C , D ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( C ) and corresponding densitometric quantification ( D ) in HT29 cells after CALB2 knockdown or negative control (NC). ( E , F ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( E ) and corresponding densitometric quantification ( F ) in SW48 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( G , H ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( G ) and corresponding densitometric quantification ( H ) in SW480 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( I – K ) RT-qPCR analysis of CALB2 ( I ), p21(CDKN1A) ( J ), and BAX ( K ) mRNA levels in HCT116 cells transfected with NC, si1-CALB2, or si1-CALB2 combined with the p53 inhibitor PFT-α (10 μM). ( L ) CCK-8 assay of HCT116 cells in the NC, si1-CALB2, and si1-CALB2+PFT-α groups over a 96 h period. ( M , N ) Representative images of scratch wound-healing assays ( M ) and quantitative analysis of relative migration rates ( N ) in HCT116 cells at 0, 24, and 48 h post-scratching. Scale bar = 250 μm. ( O – Q ) Representative double immunofluorescence staining ( O ) and quantitative analysis of fluorescence intensity ( P , Q ) for CALB2 and p-p53-Ser33 in xenograft tumor tissues from NC and OE-CALB2 groups. Scale bar = 50 μm. (n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).
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Association of CALB2 expression with p53 signaling in colorectal adenocarcinoma. ( A , B ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( A ) and corresponding densitometric quantification ( B ) in HCT116 cells following CALB2 knockdown using two independent siRNAs (si1-CALB2 and si2-CALB2) or negative control (NC). ( C , D ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( C ) and corresponding densitometric quantification ( D ) in HT29 cells after CALB2 knockdown or negative control (NC). ( E , F ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( E ) and corresponding densitometric quantification ( F ) in SW48 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( G , H ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( G ) and corresponding densitometric quantification ( H ) in SW480 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( I – K ) RT-qPCR analysis of CALB2 ( I ), p21(CDKN1A) ( J ), and BAX ( K ) mRNA levels in HCT116 cells transfected with NC, si1-CALB2, or si1-CALB2 combined with the p53 inhibitor PFT-α (10 μM). ( L ) CCK-8 assay of HCT116 cells in the NC, si1-CALB2, and si1-CALB2+PFT-α groups over a 96 h period. ( M , N ) Representative images of scratch wound-healing assays ( M ) and quantitative analysis of relative migration rates ( N ) in HCT116 cells at 0, 24, and 48 h post-scratching. Scale bar = 250 μm. ( O – Q ) Representative double immunofluorescence staining ( O ) and quantitative analysis of fluorescence intensity ( P , Q ) for CALB2 and p-p53-Ser33 in xenograft tumor tissues from NC and OE-CALB2 groups. Scale bar = 50 μm. (n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Journal: Genes

Article Title: CALB2 Expression Is Associated with Tumor Progression and Prognosis in Colorectal Adenocarcinoma

doi: 10.3390/genes17050510

Figure Lengend Snippet: Association of CALB2 expression with p53 signaling in colorectal adenocarcinoma. ( A , B ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( A ) and corresponding densitometric quantification ( B ) in HCT116 cells following CALB2 knockdown using two independent siRNAs (si1-CALB2 and si2-CALB2) or negative control (NC). ( C , D ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( C ) and corresponding densitometric quantification ( D ) in HT29 cells after CALB2 knockdown or negative control (NC). ( E , F ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( E ) and corresponding densitometric quantification ( F ) in SW48 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( G , H ) Western blot analysis of p-p53-Ser33 and total p53 protein levels ( G ) and corresponding densitometric quantification ( H ) in SW480 cells after CALB2 overexpression (OE-CALB2) or negative control (NC). ( I – K ) RT-qPCR analysis of CALB2 ( I ), p21(CDKN1A) ( J ), and BAX ( K ) mRNA levels in HCT116 cells transfected with NC, si1-CALB2, or si1-CALB2 combined with the p53 inhibitor PFT-α (10 μM). ( L ) CCK-8 assay of HCT116 cells in the NC, si1-CALB2, and si1-CALB2+PFT-α groups over a 96 h period. ( M , N ) Representative images of scratch wound-healing assays ( M ) and quantitative analysis of relative migration rates ( N ) in HCT116 cells at 0, 24, and 48 h post-scratching. Scale bar = 250 μm. ( O – Q ) Representative double immunofluorescence staining ( O ) and quantitative analysis of fluorescence intensity ( P , Q ) for CALB2 and p-p53-Ser33 in xenograft tumor tissues from NC and OE-CALB2 groups. Scale bar = 50 μm. (n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The p53-specific inhibitor PFT-α hydrobromide (PFT-α, 10 μM; MCE, Monmouth Junction, NJ, USA) was introduced to the culture medium 24 h after siRNA transfection.

Techniques: Expressing, Western Blot, Knockdown, Negative Control, Over Expression, Quantitative RT-PCR, Transfection, CCK-8 Assay, Migration, Double Immunofluorescence Staining, Fluorescence