Fig 1: KLF8 promotes the viability and migratory ability of bladder cancer cells by downregulating the expression of miR-132.a RT-qPCR was used to examine the expression of miR-132 and KLF8 in response to miR-132 mimic and oe-KLF8. b Western blot assay was used to examine the expression of KLF8 normalized to GAPDH in response to miR-132 mimic and oe-KLF8. c MTT assay was used to examine the cell viability in response to miR-132 mimic and oe-KLF8. d Flow cytometry was used to examine apoptosis in response to miR-132 mimic and oe-KLF8 under etoposide induction. e Transwell assay was used to evaluate cell migration in response to miR-132 mimic and oe-KLF8 (×200). f Western blot assay was used to measure the apoptosis-related proteins Bax and BCl-2, tumor suppressor proteins p53 and p21, and metastasis-related proteins vimentin, N-cadherin, and E-cadherin normalized to GAPDH in response to miR-132 mimic and oe-KLF8. *P < 0.05 versus the oe-NC + NC mimic group; #P < 0.05 versus the oe-KLF8 + NC mimic group. The experimental results are measurement data, and are expressed as the mean ± standard deviation. Comparisons among multiple groups were conducted by one-way ANOVA with Tukey’s post hoc test. Statistical analysis in relation to time-based measurements within each group was realized using repeated-measures ANOVA, followed by Bonferroni’s post hoc test. Cell experiments were independently repeated three times.
Fig 2: The expression of circSAMD4A, miR-218-5p and KLF8 in DXR-resistant osteosarcoma tissues and cell lines. (a–p) Analysis of circSAMD4A, miR-218-5p and KLF8 expression levels in osteosarcoma tissues and matched non-tumor tissues (a, e, i and m), osteosarcoma cell lines (HOS and U2OS) and normal cell line hFOB (b, f, j and n), treatment-resistant group and treatment-responsive group (c, g, k and o) and DXR-resistant cell lines HOS/DXR and U2OS/DXR and their parental HOS and U2OS cells (d, h, l and p) using qRT-PCR or western blot. Each experiment was repeated three times, and the average was taken. (q–s) Correlation analysis between miR-218-5p and circSAMD4A or KLF8. *P < 0.05.
Fig 3: CircSAMD4A positively regulates KLF8 via miR-218-5p. (a and b) Analysis of KLF8 expression level in HOS/DXR and U2OS/DXR cells transfected with si-NC, si-circSAMD4A, si-circSAMD4A + anti-NC or si-circSAMD4A + anti-miR-218-5p using qRT-PCR or western blot. Each experiment was repeated three times, and the average was taken. *P < 0.05.
Fig 4: NEDD4 promotes bladder cell viability and migratory ability via the NRF2/KLF8/miR-132 axis.a RT-qPCR and western blot assay was used to examine the silencing efficiency of siNRF2-1 and siNRF2-2. b RT-qPCR was used to examine the expression of NEDD4, KLF8, miR-132, and NRF2 in response to oe-NEDD4 and siNRF2. c Western blot assay was used to examine the expression of NEDD4, KLF8, and NRF2 normalized to GAPDH in response to oe-NEDD4 and siNRF2. d MTT assay was used to examine cell viability in response to oe-NEDD4 and siNRF2. e Flow cytometry was used to examine apoptosis in response to oe-NEDD4 and siNRF2 under etoposide induction. f Transwell assay was used to examine cell migration in response to oe-NEDD4 and siNRF2 (×200). g Western blot assay was used to examine expression of the apoptosis-related proteins Bax and BCl-2, tumor suppressor proteins p53 and p21, and metastasis-related proteins vimentin, N-cadherin, and E-cadherin normalized to GAPDH in response to oe-NEDD4 and siNRF2. *P < 0.05 versus the si-NC or oe-NC + si-NC group; #P < 0.05 versus the oe-NEDD4 + si-NC group. The experimental results are the measurement data and are expressed as the mean ± standard deviation. Comparisons among multiple groups were conducted by one-way ANOVA with Tukey’s post hoc test. Statistical analysis in relation to time-based measurements within each group was performed using repeated-measures ANOVA, followed by Bonferroni’s post hoc test. Cell experiments were independently repeated three times.
Fig 5: KLF8 is overexpressed in human lung cancer tissues. a The mRNA level of KLF8 is overexpressed in human lung cancer tissues (n = 34) compared with non-cancer tissues (n = 16). ***p < 0.001. b Western blot results showing KLF8 expression in human lung cancer tissues and non-cancer tissues. c Quantifications of KLF8 protein level analyzed by western blot. N = 6, **p < 0.01. d Representative histochemical staining of KLF8 expression in non-cancer and lung cancer tissues. Bar = 100 µm. e Quantifications of KLF8 expression in non-cancer (n = 5) and lung cancer tissues (n = 5).***p < 0.001. f The mRNA level of KLF8 is overexpressed in human lung cancer cell lines (A549, H1975, H1299, H460, and H520) compared with normal lung epithelial cell line (BEAS-2B). *p < 0.05, **p < 0.01 and ***p < 0.001. n = 3 in each group
Supplier Page from Abcam for Anti-KLF8 antibody