Fig 2: JunB expression during skin maturation and upon stress. a Representative microphotographs with double immunostaining for JunB (green) and FABP5 (red), indicative of sebaceous glands, in skin derived from 1, 3, and 5 days old mice. Inset showing magnified view of a sebaceous gland. Scale bars, 50 µm. b Immunostaining of JunB (green) and FABP5 (red) in 60 days old mice. Inset showing magnified view. c Representative microphotographs with double immunostaining of JunB (green) and FABP5 (red) in dorsal skin of 60 days old mice following hair plucking or d after topical TPA application, a potent inducer of proliferation. e Immunostaining of JunB (green) and LRIG1 or CD34 (red) in dorsal skin of 60 days old mice following hair plucking. f Immunostaining of JunB (green) and FABP5 (red) in adult human skin. E, epidermis; D, dermis; HF, hair follicle; SG, sebaceous gland; SD, sebaceous duct; HS, hair shaft. Asterisk indicates hair shaft autofluorescence. Dashed line indicates the epidermal-dermal junction. Scale bars, 50 µm

Fig 3: BRAF/MAPK/AP-1 axis promotes TBX3 transcription in PTC.a, b Western blot of TBX3 in PTC or normal thyroid cells (Nthy-ori-3-1) with BRAF or BRAFV600E over-expression (a) or K1 cells with BRAF knock-down (b). c K1 cells were treated with BRAF inhibitor PLX4032 or ERK1/2 inhibitor SCH772984 for 24 h, the level of TBX3 was analysis by western blot. d TBX3 and AP-1 factors levels in K1 cells over-expressing c-Jun, JunB, or c-Fos, and treated with PLX4032 or SCH772984. e TBX3 and AP-1 factors levels in K1 cells with BRAFV600E over-expression and AP-1 knock-down. f AP-1 binding motifs were analyzed through TBX3 promoter region using Jasper database. g Construction of TBX3 promoter GLuc with potential AP-1-binding sites. The predicted -358bp and -362bp site were overlapped as -360bp site. Arrows represented primers were used for qPCR. h, i Promoter GLuc activities were analyzed in HEK293T cells co-transfected with TBX3 truncated promoter (h) or TBX3-149-MUT promoter (i) and AP-1 expression constructs. j Quantitative ChIP (qChIP) analysis with anti-IgG, c-Jun, JunB and c-Fos for -149 site in K1 cells, primers targeting GAPDH gene were used as control. k Luciferase activities of GLuc-149 were analyzed in K1 cells treated with PLX4032 or SCH772984 for 12 h, 24 h or 36 h. l THCA datasets were downloaded from TCGA database to analyze the correlation between TBX3 and BRAF, n = 501. Densitometric analyses of western blot were shown (a–e). Two independent experiments were carried out with similar results for each kind of cells (a–c), and n = 3 biological independent samples (d, e, h–k). Data are shown as the mean ± s.d. (h–k). P values were calculated by unpaired two-tailed Student’s t test (h–k). Uncropped immunoblots and statistical source data are provided in Source Data.

Fig 4: Prediction and validation of the identified system by pharmacological perturbation.(A) The predictive simulation result and experimental result by PACAP stimulation in the presence (black) or absence (blue) of trametinib. Lines, simulation; dots, experimental and recovered data. Experimental and recovered data of pERK and pCREB, and the simulated data of c-Jun, c-Fos, Egr1, FosB, and JunB are given as Inputs, and simulation was performed using the NARX model in Fig 5 (see “Simulation of the integrated NARX model” section in Materials and methods). In the experiment, PC12 cells were treated in the absence (blue dots) or in the presence (black dots) of trametinib (10 μM) added at 30 min before stimulation with PACAP (100 nM). Note that the PACAP stimulation data are used, as in Fig 4. (B) Simulation using experimental and recovered data as Inputs. For each set of the Inputs (left panel for each) and Outputs (right panel for each), the unequally spaced time series data were recovered (pluses) (right panel for each), and the responses of Outputs were simulated by the NARX model identified in Fig 5A–5C (solid lines) (right panel for each).

Fig 5: Inactivation of TGF-ß signaling suppresses pancreatic ductal adenocarcinoma (PDAC)–driven ß-cell depletion.(A) Formalin-fixed paraffin-embedded (FFPE) sections from control, KC, or KSC mice were stained with H&E or subjected to immunohistochemistry analysis using antibodies to CK19 or JunB. Representative pictures of normal or PDAC tissues are shown. Bar, 200 µM. (B) FFPE sections from control, KC, or KSC mice (n = 6) were immunostained with anti-insulin antibody and revealed by immunofluorescence (IF). Then, INS+ cells in all islets from six different sections were counted, and results are presented as percentage of INS+ cells relative to the total cell number in islets. Bar, 200 µM. (C) FFPE sections from control, KC, or KSC mice (n = 6) were immunostained with anti-glucagon antibody and revealed by IF. Representative pictures of normal or remnant islets are shown (left). Percentage of remnant islets is shown. GCG+ cells in all islets from six different sections were counted, and results are presented as percentage of GCG+ relative to the total cell number in islets (middle) or as mean of the total number of GCG+ cells per islet cells (right). Bar, 200 µM. (D) FFPE sections from control, KC, or KTC mice were stained with H&E or immunostained with antibodies to CK19 or JunB and revealed by immunohistochemistry. Representative pictures of normal or PDAC tissues are shown. (E, F, G) FFPE sections from control, KC, or KTC mice (n = 6) were immunostained with antibodies to insulin or glucagon and revealed by IF. (E, F, G) Then, INS+ or GCG+ cells in all islets from six different sections were counted, and results are presented as percentage of INS+ or GCG+ cells relative to the total cell numbers in islets (E, F) or as mean of the total number of GCG+ cells per islet cells (G). For (B, C [middle], E, G), data are expressed as mean ± SEM. For (C [right], F), data are expressed as dot plot with a line at the median and whiskers showing SD. For all graphs, statistical significance was estimated by one-way ANOVA. ***P < 0.001; ns, nonsignificant.