Fig 1: Low NFIA and CRYAB expressions were associated with metastasis and poor prognosis in PCa.A NFIA and CRYAB expressions in 40 PCa tissue samples were detected by IHC. NFIA and CRYAB were spatially correlated. Representative IHC photographs of adjacent normal tissues (ANT) and tumor tissue samples (PPCa, MPCa) were shown as indicated. (i) The proportion of PCa tissues and adjacent normal tissues with negative (-), weak (+), moderate (++) and strong (+++) NFIA or CRYAB staining intensity. (ii) NFIA and CRYAB protein expressions in PCa tissues and adjacent normal tissues were quantified, and their expression were significantly decreased in PCa tissues with metastasis (iii). B NFIA and CRYAB expression levels in PCa tissue were positively correlated. C Kaplan–Meier analysis of overall survival curves of PCa patients with different NFIA or CRYAB expression levels. Magnification, ×200. Scale bars, 100 µm. The data were presented as means ± SD. nsP > 0.05; **P < 0.01; ****P < 0.0001; Student’s t-test. ANT, adjacent normal tissues; P or PPCa, primary localized PCa tissues; M or MPCa, metastatic PCa tissues; HR, hazard ratio.
Fig 2: TF-TF (bait-bait) interactions of 109 TFs studied.a Of 109 studied TFs, 80 had 202 interactions with other studied TFs. Blue edges indicate interactions from the BioID analysis, red from the AP-MS analysis and green from both. b Most of these TF-TF (118) interactions were TF interactions with NFIs (left panel). The right panel shows the separate groups shared by one or multiple NFIs. Color code: Green nodes = NFIs, yellow nodes = interactions to NFIs, orange nodes = interactions from NFIs, red nodes = interactions to and from NFIs and gray nodes = no interactions to or from NFIs. Color coding of the nodes is shown on the right side of the figure. The edge weight displays the average spectral count value detected for the corresponding interaction. c NFIA was silenced using siRNA transfection, and NFIA levels were detected 48 h after transfection by western blotting using a specific antibody against NFIA. d TF activity was investigated after NFIA silencing using both repressive and activating reporter gene analysis. Both the repressing and activating functions of KLF4 were reduced upon NFIA silencing. In addition, SOX2 and PAX6 activity was reduced, while HME1 activity was increased upon NFIA silencing. The representative data from two repeated experiments are presented as mean values +SD as appropriate. Two-sided t-test was used. N = 3, ***p < 0.001, **p < 0.01, *p < 0.05. Source data of the figure are provided as a Source Data file.
Fig 3: NFIA was the direct target of miR-671.A, B Identify potential miR-671 target genes by 4 common miRNA prediction databases and GSE21034. C–E The correlations between miR-671 expression and CFL2, NFIA, and RBMS3 expression. F, G CFL2, NFIA and RBMS3 mRNA and protein expressions in RWPE-1 and PCa cell lines were examined by qPCR and WB. H WB analysis revealed a negatively correlation of NFIA expression with miR-671 expression in PCa cells. RBMS3 did not accept the regulation of miR-671. I NFIA IHC staining in xenografts with LV-in-miR-671 cells or LV-in-NC cells. J-Left: potential miR-671 binding sequences in the 3’UTR of NFIA mRNAs. J-Right: luciferase activity assays showed that the overexpression of miR-671 significantly reduced the luciferase activity of binding site of NFIA, and the mutation of binding site blocked the interaction. Magnification, ×200. Scale bars, 100 µm. The data were presented as means ± SD from three biological replicates. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; Student’s t-test. N, adjacent normal tissues; P, primary localized PCa tissues; M, metastatic PCa tissues; WT, wide type; MT, mutant type.
Fig 4: miR-181d-5p relieves astrocyte development, oxidative stress, and inflammation by targeting NFIA. (a) GFAP staining (scale bar = 40 µm). (b) SOD activity. (B) MDA content. (d) Western blot of Bcl-2 and Bax protein. (e) Proinflammatory factor levels were measured by ELISA. **P < 0.01 and ***P < 0.001 vs. the sham group; #P < 0.05, ##P < 0.01, and ###P < 0.001 vs. the model group; &P < 0.05, &&P < 0.01, and &&&P < 0.001 vs. the miR-181 group.
Fig 5: miR-181d-5p directly targeted the NFIA gene. (a) Representative western blot of nuclear factor I-A (NFIA). **P < 0.01 vs. the sham group. (b) The binding sites of miR-181d-5p in the NFIA sequence by starBase prediction. (c) Expression of miR-181d-5p by RT-qPCR detection. ***P < 0.001. (d) The interaction between the NFIA 3'UTR and miR-181d-5p was analyzed by a dual-luciferase reporter gene. 293T cells were cotransfected with miR-181d-5p mimic and WT-NFIA or miR-181d-5p mimic and MUT-NFIA. ***P < 0.001. (e) Representative western blot of NFIA. ***P < 0.001.
Supplier Page from Abcam for Anti-CTF/NFIA antibody