EMBO/soda-nel huggingface dataset:
VEGF blockade enhances the antitumour effect of BRAFV660E inhibition (2016)
Figure 1

Figure 1. Effect of the combination of PLX4720 and bevacizumab on the growth of A375, COLO205 and MC-1 cells harboring BRAFV600E transplanted in athymic nude mice.

(A) Mice bearing established A375 were treated with vehicle (n=7), PLX4720 (n=5) bevacizumab (n=7) or COMBO (n=5). Tumor growth is expressed as % change of the initial tumor, **P < 0.01, ***P < 0.001 versus vehicle (PLX4720 P = 0.0049; bevacizumab P = 0.0003; P = 4.86E-10) ŦP < 0.05 compared to PLX4720 (P = 0.015).

(B) Mice bearing established COLO205 were treated with vehicle (n=7), PLX4720 (n=8) bevacizumab (n=7) or COMBO (n=8). Tumor growth is expressed as % change of the initial tumor, ***P < 0.001 versus vehicle (PLX4720 P = 2.75E-05; bevacizumab P = 2.96E-05; COMBO P = 2.13E-07) Ŧp < 0.05 compared to PLX4720 (P = 0.039).

(C) Melanoma MC-1 cells (5 x 105) were injected into the tail vein of CD1 mice. Lung colonization was assayed by HE staining and calculating the number of nodules and their total area normalized per the total area of the lungs. The mice analyzed were: start point, n=6; vehicle, n=7; PLX4720, n=6; bevacizumab, n=6; COMBO, n=7, *P < 0.001 versus vehicle (P = 0.031).

(D) Representative images of tumor cell proliferation determined by immunofluorescence Ki67 staining in A375 xenografts treated as indicated. Bar graphs indicate the Ki67+ area/tumor area (n=4 tumors), ***P < 0.001 versus vehicle (PLX4720 P = 2.83E-08; bevacizumab P = 4.25E-13; COMBO P = 6.16E-08).

(E) Representative images of tumor cell apoptosis determined by immunofluorescence staining with TUNEL in A375 xenografts treated as indicated. Bar graphs indicate the TUNEL+ area/tumor area (n=3 tumors). ***P < 0.001 versus vehicle (P = 6.2.7E-10).

The scale bars represent 1 cm (A-B), 100 µm (D) and 50 µm (E). The results are given as the mean ± SEM. Significance was assessed by one-way ANOVA test followed by post-hoc pairwise analysis test (A-E).
VEGF blockade enhances the antitumour effect of BRAFV660E inhibition (2016)
Figure 2

Figure 2. Vascular response of A375 and COLO205 xenograft tumors to PLX4720 and bevacizumab.

(A) Representative images of vasculature stained by an antibody anti-CD31 in A375 xenografts treated as indicated. Bar graphs indicate quantitative micro vessel density (MVD) and micro vessel area (MVA) analysis (n=5 tumors), ***P < 0.001 versus vehicle (MVD: bevacizumab P = 5.94E-25; COMBO P = 1.84E-04) (MVA: PLX4720 P = 4.98E-09; bevacizumab P = 1.16E-19; COMBO P = 3.00E-08).

(B) Representative images of vessel lumen in A375 xenografts treated as indicated. Bar graphs indicate the quantitative analysis of lumen diameters in A375 and COLO205 xenografts (n=3 tumors), *P < 0.05, **P < 0.01, ***P < 0.001 versus vehicle (A375: PLX4720 P = 0.0078; COMBO P = 0.0012) (COLO205: PLX4720 P = 2.01E-06; COMBO P = 1.33E-07).

(C) Representative images of perfusing fluorescent beads and their relationship with microvessels in A375 xenografts treated as indicated. Bar graphs indicate the % of CD31+ vessels co-stained with fluorescent beads (n=4 tumors), **P < 0.01 versus vehicle (PLX4720 P = 0.0031; COMBO P = 0.0033).

(D) Representative images of hypoxia marker pimonidazole in A375 xenografts treated as indicated. Bar graphs show the % of tumor hypoxic area in A375 and COLO205 xenografts (n=5 tumors), **P < 0.01, ***P < 0.001 versus vehicle (A375: PLX4720 P = 6.94E-12; COMBO P = 3.34E-12) (COLO205: PLX4720 P = 0.0015; bevacizumab P = 0.0004; COMBO P = 0.0003).

The scale bars represent 100 µm in (A-D) and 50 µm in (B-C). The results are given as the mean ± SEM. Significance was assessed by one-way ANOVA test followed by post-hoc pairwise analysis test.
VEGF blockade enhances the antitumour effect of BRAFV660E inhibition (2016)
Figure 4

Figure 4. COMBO treatment increases macrophage infiltration

(A) Representative images of leukocytes infiltration determined by immunofluorescence CD45 staining in of A375 xenografts treated as indicated. Bar graphs indicate the CD45+ area/tumor area in A375 and COLO205 xenografts (n=5 tumors), ***P < 0.001 versus vehicle (A375: P = 2.19E-20) (COLO205: P = 1.81E-06).

(B) Representative images of macrophage infiltration determined by F4/80 immunofluorescence staining in A375 xenografts treated as indicated. Bar graphs indicate the F4/80+ area/tumor area in A375 and COLO205 xenografts (n=5 tumors) ***P < 0.001 versus vehicle (A375: COMBO P = 3.42E-13) (COLO205: COMBO P = 1.94E-06).

(C) A375 xenograft tumors after 14 days of treatment were subjected to FACS and tumor infiltrate was analyzed. Gating strategy for CD45+ cells and graph showing the quantification of FACS analysis of infiltrating CD45+ leukocytes in vehicle (n=4 tumors) compared to COMBO (n=6 tumors) treated tumors, **P < 0.01 versus vehicle (P = 0.0064).

(D) Graph showing the quantification of FACS analysis of infiltrating F4/80+ macrophages and CD11c+MHCII+ dendritic cells in vehicle (n=4 tumors) compared to COMBO (n=6 tumors) treated tumors, **P < 0.01 versus vehicle (TAM P = 0.0061; DC P = 0.0091).

The scale bars represent 100 µm in (A-B). The results are given as the mean ± SEM. Significance was assessed by one-way ANOVA test followed by post-hoc pairwise analysis test (A-B) and Student’s t-test (C-D).
VEGF blockade enhances the antitumour effect of BRAFV660E inhibition (2016)
Figure 5

Figure 5. Macrophages infiltrated after COMBO treatment are polarized toward M1-like phenotype

(A) Real-rime quantitative PCR of the indicated genes (M1-like and M2-like macrophages markers) of A375 xenograft treated with PLX4720, bevacizumab or COMBO. Data are presented as expression fold change (log2) compared with vehicle after normalization for housekeeping gene TBP (n=3 tumors), (*P < 0.05, **P < 0.01, ***P < 0.001 versus vehicle (PLX4720: m-Arg1 P = 5.65E-06) (bevacizumab: m-Ccl5 P = 0.025) (COMBO: m-Ccl5 P = 0.0070; m-Cd40 P = 0.00099; m-Cxcl10 P = 0.043; m-Cxcl9 P = 0.046; m-Il1b P = 0.0367; m-Stat1 P = 0.049; m-Tlr2 = 0.00182; m-Arg-1 P = 0.0013).

(B) Representative images of COMBO-treated tumor sections co-stained with an antibody against CD68 and an antibody anti-CCR7 (M1-like polarization) or anti-C206 (M2-like polarization).

(C) Graphs showing the quantification of FACS analysis of infiltrating F4/80+ tumor macrophages and average percentage of median fluorescence intensity (MFI) of TNFα+ or INOS+ on F4/80+ cells after ex vivo stimulation with PMA and ionomycin in vehicle (n=4 tumors) and COMBO (n=6 tumors), *P < 0.05 versus vehicle (TNFα P = 0.037; INOS = 0.038).

(D) Graphs showing the quantification of FACS analysis of infiltrating CD11+Ly6C+ tumor monocytes and average percentage of MFI of TNFα+ or INOS+ on CD11+Ly6C+ cells after ex vivo stimulation with PMA and ionomycin in vehicle (n=4 tumors) and COMBO (n=6 tumors), *P < 0.05 versus vehicle (TNFα P = 0.048; INOS = 0.038).

(E) Graph showing the quantification of circulating CD11+Ly6C+ monocytes in peripheral blood analyzed by FACS in vehicle- (n=4 tumors) and COMBO- (n=6 tumors) treated tumors.

(F) Cell viability measured as fluorescence intensity of ZsGreen-A375 cells co-cultured with cells isolated from xenograft treated with vehicle or COMBO with after 48h. Data are representative of one experiment out of two done in triplicate *P < 0.05 versus vehicle (P = 0.046).

The scale bars represent 50 µm in (B). The results are given as the mean ± SEM. Significance was assessed by Student’s t-test (A-F).
VEGF blockade enhances the antitumour effect of BRAFV660E inhibition (2016)
Figure 6

Figure 6. COMBO treatment reduces collagen deposition in A375 xenografts

(A) Representative images of collagen deposition determined by collagen I immunofluorescent staining in A375 xenografts treated as indicated. Bar graphs indicate the collagen I+ area/tumor area (n=5 tumors), ***P < 0.001 versus vehicle (P = 2.93E-06).

(B) Representative images of CAF density determined by -SMA immunofluorescence staining in A375 xenografts treated as indicated. Bar graphs indicate -SMA+ area/tumor area (n=5 tumors), ***P < 0.001 versus vehicle (P = 0.0006).

(C) Representative images of Lysyl Oxidase enzime determined by LOX immunofluorescence staining in A375 xenografts treated as indicated. Bar graphs indicate the LOX+ area/tumor area (n=3 tumors), *P < 0.05 versus vehicle (bevacizumab P = 0.033; COMBO P = 0.040).

(D) Ingenuity Pathway Analysis (IPA) identified TGFβ1 as one of the most significant upstream regulators (activation z-score=-4.753, P = 2.1E10E-29; predicted activation state: inhibited) of the specifically modulated genes after COMBO treatment, according to microarray analysis. Shades of red indicate the degree of up- regulation while shades of green indicate the degree of down-regulation of significant downstream genes.

(E) Quantitative Real-rime PCR of h-TGFB1 and m-Tgfb1 of A375 xenograft treated with PLX4720, bevacizumab or COMBO. Data are presented as expression fold change (log2) compared with vehicle after normalization for the housekeeping gene TBP (n=3 tumors), *P < 0.05 versus vehicle (P = 0.043).

(F) Western blot for human TGFβ in total protein extract from A375 tumors treated as indicate. Vinculin was used as an internal control.

The scale bars represent 100 µm in (A-C). The results are given as the mean ± SEM. Significance was assessed by one-way ANOVA test followed by post-hoc pairwise analysis test (A-C) and Student’s t-test (E-F).
VEGF blockade enhances the antitumour effect of BRAFV660E inhibition (2016)
Figure 7

Figure 7. Bevacizumab delays tumor relapse after acquired resistance to PLX4720 in A375 xenografts through macrophage recruitment.

(A) Effect of clodronate liposomes on tumor growth inhibition induced by COMBO treatment in in mice bearing A375 tumors. Tumor growth is expressed as % change of the initial tumor. Macrophage depletion by clodronate liposome enhanced the tumor growth of tumors treated with the combination of PLX4720 and bevacizumab (n=5 mice/group), *P < 0.05, ***P < 0.001 versus vehicle (vehicle Clodronate P = 0.049, COMBO P = 5.72E-06) ŦP < 0.05 compared to COMBO (P = 0.033).

(B) Representative images of macrophage infiltration determined by F4/80 immunofluorescence staining in A375 xenografts treated as indicated. Bar graphs indicate the F4/80+ area/tumor (n=3 tumors), ***P < 0.001 versus vehicle (P = 1.53E-25).

(C) Representative images of collagen deposition determined by collagen I immunofluorescence staining in A375 xenografts treated as indicated. Bar graphs indicate the collagen I+ area/tumor (n=3 tumors).

(D) Waterfall plots showing the percent change in volume (relative to the initial tumor volume) for the individual A375 xenografts in each treatment group (vehicle, PLX4720, bevacizumab and COMBO) from week 1 to week 6.

(E) Representative images of macrophage infiltration determined by F4/80 immunofluorescence staining in a responder and a relapsing A375 xenografts. Bar graphs indicate the F4/80+ area/tumor (n=3 mice), ***P < 0.001 versus vehicle (P = 1.25E-06).

(F) Representative images of collagen deposition determined by collagen I immunofluorescence staining in a responder and a relapsing A375 xenografts. Bar graphs indicate the collagen I+ area/tumor (n=3 mice), ***P < 0.001 versus vehicle (P = 4.03E-05).

(G) Real-rime quantitative PCR of h-TGFB1 of A375 xenograft treated with PLX4720, bevacizumab or COMBO. Data are presented as expression fold change (log2) of relapsing tumors compared to responder tumors after normalization for housekeeping gene TBP (n=3 tumors) ***P < 0.001 versus vehicle (P = 0.0006).

The scale bars represent 100 µm (B-C-D-F). The results are given as the mean ± SEM. Significance was assessed by one-way ANOVA test followed by post-hoc pairwise analysis test (A) and and Student’s t-test (B-C-E-F-G).
Neuronal carbonic anhydrase VII provides GABAergic excitatory drive to exacerbate febrile seizures. (2013)
Figure 1

Generation of CA VII-deficient mice. (A) Targeting strategy for the Car7 locus. Endogenous Car7 locus (top). Targeting construct (middle). Targeted locus after Cre-mediated recombination (bottom). Exons are indicated as grey boxes and loxP sites as arrowheads. (B) Multiple tissue northern blot for CA VII. In the P56 mouse, CA VII is prominently expressed in the central nervous system including brain and spinal cord. Gapdh served as the loading control. (C) Developmental profile of CA VII protein expression. Western blot analysis of protein lysates of whole hippocampi shows increasing CA VII levels from P3 to P30. The band of the appropriate size (∼30 kDa) was absent in protein lysates from KO tissue. (D) In western blots from protein lysates of cultured cells, CA VII was detected in mixed glia/neuron cultures but not in glia cell cultures. The presence of astrocytes in all cultures was confirmed by detection of GFAP. Actin served as the loading control in C and D.
Neuronal carbonic anhydrase VII provides GABAergic excitatory drive to exacerbate febrile seizures. (2013)
Figure 2

Development of cytosolic CA activity in mouse CA1 pyramidal neurons is based on sequential expression of isoforms VII and II. (A) Original single-cell pHi traces and their time derivatives from P5 and P14 WT and P14 CA VII KO neurons (baseline pHi 7.21, 7.13 and 7.11, respectively). Superfusion with CO2/HCO3−-free HEPES solution (upper horizontal bars) evoked an intracellular alkalinization, which in P14 WT was large and suppressed by 100 μM acetazolamide (AZ, lower horizontal bar), indicating the presence of CA activity (see Supplementary Materials and methods). The possible effect of AZ on extracellular CAs was excluded by adding 10 μM benzolamide (a poorly permeant CA inhibitor). Inset shows an overlay of the BCECF fluorescence signal and Dodt gradient contrast image of CA1 pyramidal neurons in P14 WT (scale bar 10 μm). (B) Summary of the results obtained using the cytosolic CA activity detection method shown in A and quantified as the percentage of cells showing cytosolic CA activity. Data from CA II KO and CA II/VII KO neurons were obtained only at >P35. The number of cells tested is indicated for each bar. The animal numbers for WT mice at the different age points was n=3 (P5–8), n=2 (P10–11), n=3 (P14–16), n=2 (P17–18), n=2 (P20–30) and n=2 (>P35); for CA VII KO mice the numbers were n=3 (P5–8), n=2 (P10–11), n=5 (P14–16), n=3 (P17–18), n=4 (P20–30) and n=2 (>P35); and for CA II KO and CA II/CA VII KO mice the number was n=2 (>P35). (C) Typical single pyramidal neuron pHi responses evoked by withdrawal of CO2/HCO3− (horizontal bar, HEPES) in slices from >P35 WT, CA VII KO and CA II/VII KO mice. The rate of rise and the amplitude of the alkaline shift are identical in WT (P39, baseline pHi 7.01) and CA VII KO neurons (P40, 7.06), whereas alkalinization develops much more slowly in the CA II/VII KO (P46, 7.04).
Neuronal carbonic anhydrase VII provides GABAergic excitatory drive to exacerbate febrile seizures. (2013)
Figure 3

CA VII and CA II similarly enhance GABAAR-mediated depolarization in CA1 pyramidal neurons. (A) Electrical stimulation (40 pulses, 100 Hz at stratum radiatum/stratum lacunosum-moleculare (sr/slm) border (horizontal bar)) in the presence of AP5/CNQX/CGP55845) evokes an excitatory GABAergic response in WT, but not in CA VII KO neurons. Picrotoxin blocked the biphasic response (PiTX 90 μM; n=3/genotype). The mean initial slope of the depolarizing shift was faster in WT than in CA VII KO (13.53±1.27 versus 6.39±0.96 mV/s, P<0.001, Student's t-test). The resting membrane potential of WT and CA VII KO neurons did not differ (−71.0±1.1 versus −72.3±1.4 mV, respectively; n=15 neurons for both genotypes, P=0.47, Student's t-test). (B) Microinjection of GABA (5 mM for 100 ms, sr/slm border in the presence of CGP55845/TTX, arrowheads) induced a pronounced depolarization in WT neurons (black) that was abolished in the absence of CO2/HCO3− (grey). The depolarization was smaller in CA VII KO (red). Bar diagram illustrates GABAergic mean peak depolarization in P12–16 WT and CA VII KO (13.3±0.5 versus 8.3±0.6 mV, P<0.001, Student's t-test). (C) In the presence of AP5/CNQX/CGP55845, microinjection of GABA (5 mM, sr/slm border) triggered SR 95531-sensitive field potential spikes in WT slices already upon 8-ms injection, while even 16-ms injection evoked hardly any spikes in CA VII KO (left). Mean number of spikes was plotted against GABA puff duration in P12–16 WT and CA VII KO (P<0.01 and P<0.001, Student's t-test; right). (D) After P35, depolarizations upon GABA microinjection (arrowheads) in the presence of CGP55845/TTX were indistinguishable in WT (black), CA VII KO (red) and CA II KO (blue) but smaller in CA II/VII KO (green). The depolarization in WT was abolished in the absence of CO2/HCO3− (grey, n=5). Bar diagram summarizes mean peak depolarization for each genotype (right; ANOVA, Bonferroni). *P<0.05, **P<0.01 and ***P<0.001. Error bars denote s.e.m. and n is the number of slices (C) or cells tested (one cell per slice) (A, B and D).
Neuronal carbonic anhydrase VII provides GABAergic excitatory drive to exacerbate febrile seizures. (2013)
Figure 4

CA VII promotes experimental febrile seizures in P13–14 mice. (A) Representative raw traces of epidural EEG recordings during hyperthermia (HT) from a WT and a CA VII KO mouse. Seizures were reliably induced in WT (n=9) but not in CA VII KO (n=7). (B) Specimen recordings of breath rate from a WT and a CA VII KO mouse. The baseline breath rate (WT 153±11 breaths/min and CA VII KO 157±8 breaths/min) was increased during HT in both genotypes (WT 272±28 breaths/min and CA VII KO 279±12 breaths/min). The bar diagram summarizes the mean increase in breath rate in WT (177±12%) and CA VII KO mice (178±6%, P=0.98). (C) Hyperthermia caused a comparable alkalosis in blood pH in both WT and CA VII KO mice. The blood pH did not differ between the two genotypes under control conditions (P=0.37) or during HT (P=0.44). (D) Electrographic seizures were not detected in WT mice when hyperthermia was induced in the continuous presence of 5% CO2 (n=5). Error bars denote s.e.m., all P-values are based on Student's t-test (***P<0.001), and animal number is given in bar diagrams. Rectal temperatures are given above the traces.
Neuronal carbonic anhydrase VII provides GABAergic excitatory drive to exacerbate febrile seizures. (2013)
Figure 6

CA VII expression during human brain development. Line plots show the log2-transformed exon array signal intensity (left), and the normalized ratio of absolute copy number of CA VII to Gapdh by droplet digital PCR (ddPCR, right) in the neocortex (NCX) and hippocampus (HIP) from the early fetal period to late adulthood. The solid line between periods 7 and 8 separates the prenatal from the postnatal period.
C9orf72 ALS/FTD dipeptide repeat protein levels are reduced by small molecules that inhibit PKA or enhance protein degradation (2021)
Figure 3

Figure 3. GELD and SPL enhance the degradation of DPRs.

A, B Lysates from NSC34 cells treated for 24 h and immunoblotted for SQSTM1/p62 and LC3 expression. (B) Quantification from three biological replicates. Data are mean ± SEM. Two-tailed, unpaired t-test, **P = 0.0047, *P = 0.0393.

C Relative expression level of Sqstm1/p62, Lc3, Tfeb, Hspb8 and Bag3 mRNAs in NSC34 cells treated for 24 h. Data are mean ± SEM from three biological replicates. 2-way ANOVA followed by Dunnett's multiple comparisons test, ***P<0.001. Dotted lines represent RNA expression in the DMSO control.

D Lysates from NSC34 cells transfected with a proteasome activity reporter (GFPu), treated for 24 h and immunoblotted for GFP expression. Quantification from three biological replicates. Data are mean ± SEM. Two-tailed, unpaired t-test, *P = 0.0398 GELD vs DMSO, *P = 0.0124 SPL vs DMSO, *P = 0.0253 FSK vs DMSO.

E Co-immunoprecipitation (IP) analysis on 66R-derived poly-GP and ubiquitin interaction. Lysates from NSC34 cells transfected with 66R and treated with 10 µM MG132 for 16 h, immunoprecipitated for poly-GP and immunoblotted for ubiquitin and poly-GP.

F Lysates from NSC34 cells transfected with scrambled-GPx100, treated for 24 h with GELD, SPL and/or 16 h with MG132 and immunoblotted for scrambled-GP expression. Quantification from three biological replicates. Data are mean ± SEM. 1-way ANOVA followed by Uncorrected Fisher's LSD, P* = 0.0186 for GELD vs DMSO, ***P = 0.0001 GELD + MG132 vs GELD and SPL vs DMSO.

G Lysates from NSC34 cells transfected with scrambled-GPx100, treated for 24 h with SPL and/or 10 mM 3MA and immunoblotted for scrambled-GP expression. Quantification from three biological replicates. Data are mean ± SEM. Two-tailed, unpaired t-test, ****P<0.0001 for SPL vs DMSO and 3MA vs DMSO, ***P = 0.0008 SPL + 3MA vs SPL.

Concentration used: 1 μM GELD, 10 μM ERY, SPL, FSK.
C9orf72 ALS/FTD dipeptide repeat protein levels are reduced by small molecules that inhibit PKA or enhance protein degradation (2021)
Figure 5

Figure 5. RNA interference of PKA catalytic subunit β reduces DPR levels by repressing (G4C2)n translation.

A, B Lysates from HEK293T cells, transfected with RNAi scrambled or RNAi PRKACA (72 h) and 2R or 66R (24 h), immunoblotted for PKA-Cα and poly-GA (HA-tag) expression. α-Tubulin used as loading control. (B) Quantification of PKA-Cα and poly-GA expression from three biological replicates. Data are mean ± SEM. Two-tailed, unpaired t-test. PKA-Cα: ns siScr 66R vs siScr 2R, ***P = 0.0002 siPRKACA 66R vs siScr 2R, ***P = 0.0003 siPRKACA 66R vs siScr 66R. poly-GA expression: ****P<0.0001 siScr 66R vs siScr 2R, ****P<0.0001 siPRKACA 66R vs siScr 2R, **P = 0.0062 siPRKACA 66R vs siScr 66R .

C, D Lysates from HEK293T cells, transfected with RNAi scrambled or RNAi PRKACB (72 h) and 2R or 66R (24 h), immunoblotted PKA-Cβ and poly-GA (HA tag) expression. α-Tubulin used as loading control. (D) Quantification of PKA-Cβ and poly-GA expression from three biological replicates. Data are mean ± SEM. Two-tailed, unpaired t-test. PKA-Cβ: ns siScr 66R vs siScr 2R, **P = 0.0049 siPRKACB 66R vs siScr 2R, **P = 0.0091 siPRKACB 66R vs siScr 66R. poly-GA expression: ****P<0.0001 siScr 66R vs siScr 2R, ****P<0.0001 siPRKACB 66R vs siScr 2R, ***P = 0.0007 siPRKACB 66R vs siScr 66R.

E Co-sedimentation profile of (G4C2) (upper panel) and GAPDH (lower panel) mRNAs along the sucrose gradient fractions of polysomal profiles from control (grey lines) and PRKACA-depleted (orange lines) cells. Data are mean ± SEM from three independent biological replicates. One-tailed t-test, *P<0.05.

F Co-sedimentation profile of (G4C2) (upper panel) and GAPDH (lower panel) mRNAs along the sucrose fractions of polysomal profiling from control (grey lines) and PRKACB-depleted (red lines) cells. Data are mean ± SEM from three independent biological replicates. One-tailed t-test, *P<0.05; **P<0.01.

ns = not significant