Fig 2: The target CFL1 and protein quantifications were validated.(A) The dynamic results revealed that CFL1 was significantly upregulated in angiotensin-(1-7)-treated HAECs and attenuated by the Mas-receptor antagonist A779. (B) The expression of Cofilin-1 increased significantly in the groups treated with angiotensin for 6 h or 24 h and was also attenuated by A779 pre-treatment. The data are expressed as the mean ± SEM for three independent experiments. *p < 0.05 indicates a significant difference relative to the control (n = 5).

Fig 3: Atypical PKCs control actin cytoskeleton via Cofilin activation. The Cofilin pathway is activated by atypical PKC via Slingshot, SSH. Phosphorylated Cofilin at serine-3 is inactive. Phosphatases such as SSH induces the dephosphorylation and activation of Cofilin. Activated Cofilin can then sever the mother filaments and produced free barbed ends leading to the elongation of newly formed actin filaments which are preferred for dendritic nucleation by ARP2/3 complex and globular actin. These newly formed structures push on the cell membrane to promote protrusion and motility.

Fig 4: Atypical PKC controls actin organization by regulating Cofilin via SSH2 in CRC cells. (a) Immunoblot analysis is showing the effect of ICA-I and ?-Stat in healthy and CRC cells following three days of treatment. Proteins tested were pSSH (S978), SSH1, SSH2, pCofilin (S3) and Cofilin. (c) The expression of PKC-?, PKC-?, SSH1, SSH2, pCofilin (S3) and Cofilin in siPRKCI and siPRKCZ transfected CRC cells. (B & D) The bar charts are representing the corresponding band intensities. a-tubulin was probed as a loading control. All experiments were performed for at least 3 independent times. Mean ± S.E.M and * represents p value (* < 0.05, ** < 0.02, *** < 0.01).

Fig 5: Molecular validation of the RNA sequencing cutoffs.Changes in mRNA (a-f) or protein (g-h) contents of selected cerebellar dysregulated genes after normalization. a, efnb3, ephrinB3 (RNAseq rank #26/1192 in males; up-regulated; q = 0.018; p = 3.41E-05). b, sbf1, SET Binding Factor 1 (RNAseq rank #82/1192 in males; down-regulated; q = 0.067; p = 0.00039). c, fabp7, Fatty Acid Binding Protein 7 (RNAseq rank #84/1192 in males; up-regulated; q = 0.0725; p = 0.00043). d, tcf7l2, transcription factor 7 like 2 (RNAseq rank #795/1192; up-regulated; q = 0.514; p = 0.029). e, nf1, Neurofibromin 1 (RNAseq rank #1189/1192 in males; down-regulated; q = 0.59; p = 0.0499). f, plp1, Proteolipid Protein 1 (RNAseq rank #64/389 in females; down-regulated; q = 1; p = 0.0054). Normalized qRT-PCR data (to pgk1) is presented as mean fold changes ± SEM, n = 10 samples/group.). Two-way ANOVA with Sidak’s multiple comparison test (a-f). *p < 0.05, **p < 0.01, ****p < 0.0001. g-h, Cerebellum protein contents determined by Western blot in males, and normalized to GAPDH levels. hnRNP?/Cfl1:n = 7 C and 7plKO; PSAP: n = 6 C and 6 plKO. Multiple unpaired t tests with Holm-Sidak multiple comparison test (**p < 0.01, ***p < 0.005, ****p < 0.0001). PSAP, prosaposin (RNAseq rank #950/1192 in males; up-regulated; q = 0.5329; p = 0.0359). Cfl1, cofilin-1 (RNAseq rank #27/1192 in males; up-regulated; q = 0.0192; p = 4.03E-05). hnRNP?, heterogeneous nuclear ribonucleoprotein ? (rank #787; up-regulated; q = 0.509; p = 0.0284). i, mbp, myelin basic protein. Normalized qRT-PCR data (to ywhaz, Tyrosine 3-Monooxygenase/Tryptophan 5-Monooxygenase Activation Protein Zeta). Two-way ANOVA with Sidak’s multiple comparison test (n = 11 C and 11 plKO; *p < 0.05). See Extended Data Table 1 sheets 1 and 2 for list of ranked DEGs. Source data