transcription factor 6 atf 6  (Novus Biologicals)

Journal: EMBO Molecular Medicine

Article Title: Encephalopathy-linked UFM1 variants impede neuronal protein translation, development, and function

doi: 10.1038/s44321-026-00389-6

Figure Lengend Snippet: ( A ) Representative images of HPG/Alexa-488 labeled primary hippocampal UFM1-cKO neurons (black) infected at DIV 1 with viruses expressing either RFP, CRE, CRE combined with UFM1-WT (WT) or UFM1-R81C (R81C) and processed for FUNCAT analysis at DIV 10. CHX, Cycloheximide; Adj., adjusted displayed intensity. Scale bar, 50 µm. ( B ) Bar graph depicting the quantification of the HPG/Alexa-488 signal intensity as shown in black in ( A ), expressed as a percentage of the HPG/Alexa-488 signal intensity of RFP control neurons. Data were obtained from N = 3 experiments, n = 67 RFP, 60 CRE, 38 WT, 40 R81C, and 24 RFP + CHX neurons. ****p RFP-CRE = 0.000008327, ****p CRE-WT = 0.0000000001 , ****p WT-RFP+CHX = 0.0000000131, ***p RFP-RFP+CHX = 0.0001, *p CRE-RC = 0.0246, **p WT-RC = 0.0046, *p RC-RFP+CHX = 0.0228 using a D’Agostino–Pearson normality test and Kruskal–Wallis test. Bar graphs show averages ± SEM. ( C ) Anti-ATF6, PERK, and IRE1α analysis of lysates from DIV 13 UFM1-cKO primary hippocampal neurons infected at DIV 1 with viruses expressing either RFP, CRE, CRE combined with UFM1-WT (WT) or UFM1-R81C (R81C). Molecular weights are indicated on the left (kDa). ( D ) Bar graph depicting the quantification of cleaved ATF6 IB signal as shown in ( C ). N = 4 experiments with n = 3. Data were analyzed using a D’Agostino–Pearson normality test and Kruskal–Wallis test. Bar graphs show averages ± SEM. ( E ) Bar graph depicting the quantification of IRE1α IB signal as shown in ( C ). N = 4 experiments, n = 6. Data were analyzed using a D’Agostino–Pearson normality test and Kruskal–Wallis test. Bar graphs show averages ± SEM. ( F ) Bar graph depicting the quantification of PERK IB signal as shown in ( C ). N = 4 experiments, n = 6. **p RFP-CRE = 0.0079, **p CRE-WT = 0.0017 using a D’Agostino–Pearson normality test and Ordinary one-way ANOVA. Bar graphs show averages ± SEM. ( G ) Representative images of DIV 5 primary hippocampal UFM1-cKO neurons infected at DIV 1 with viruses expressing either RFP, CRE, CRE combined with UFM1-WT (WT) or UFM1-R81C (R81C) and treated with thapsigargin (Tg, 3 µM, 15 min, bottom panels) or DMSO (top panels) as control. Neurons were fixed at DIV 5 and immunolabelled with anti-KDEL antibody (black). Scale bar, 50 µm. Data were obtained from N = 3 experiments, n = 38 RFP, 28 CRE, 32 UFM1-WT (WT), 34 UFM1-R81C (R81C), 23 RFP Tg, 34 CRE Tg, 33 UFM1-WT Tg (WT Tg), and 31 UFM1-R81C Tg (R81C Tg) neurons. ( H ) Bar graph depicting the quantification of the KDEL signal intensity in control and Tg-treated neurons, as shown in ( G ), normalized against the anti-KDEL signal intensity of RFP and expressed as a percentage. Data were obtained from N = 3 experiments, n = 38 RFP, 28 CRE, 32 UFM1-WT (WT), 34 UFM1-R81C (R81C), 23 RFP Tg, 34 CRE Tg, 33 UFM1-WT Tg (WT Tg), and 31 UFM1-R81C Tg (R81C Tg) neurons. ****p RFP-RC Tg = 0.0000000041; ****p WT-RC Tg = 0.0000000016; ****p RC-RC Tg = 0.0000002162; ***p WT-WT Tg = 0.0004; **p RFP-RFP Tg = 0.0071; **p RFP-WT Tg = 0.0011; **p CRE-RC Tg = 0.0046; **p WT-RFP Tg = 0.0027; **p CRE Tg-RC Tg = 0.0044; *p RC-RFP Tg = 0.0464; *p RC-WT Tg = 0.0117 using a D’Agostino–Pearson normality test and Kruskal–Wallis test. Bar graphs show averages ± SEM. .

Article Snippet: ATF6 (1:1000 WB) , Novus Biologicals , NBP1-40256.

Techniques: Labeling, Infection, Expressing, Control